Novel 7-dehydrocholesterol derivatives and methods using same

ABSTRACT

The present invention provides, in certain aspects, novel 7-dehydrocholesterol (7DHC) derivatives that are useful in treating or preventing cancer, as well as in treating or preventing uncontrolled angiogenesis, in a subject. In certain embodiments of the present invention, the subject is a human. In other aspects, the present invention provides a method of preparing compounds of the invention, or a salt or solvate thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims priority under 35 U.S.C. § 119(e) to U.S.Provisional Application No. 61/976,148, filed Apr. 7, 2014, which ishereby incorporated by reference in its entirety herein.

BACKGROUND OF THE INVENTION

Cancer, known medically as a malignant neoplasm, refers to a broad groupof diseases involving unregulated cell growth. In cancer, cells divideand grow uncontrollably, forming malignant tumors and invading nearbyparts of the body. The cancer may also spread to more distant parts ofthe body through the lymphatic system or bloodstream, through amechanism generally known as metastasis. Not all tumors are cancerous,however. In contrast to cancerous tumors, benign tumors do not invadeneighboring tissues and do not spread throughout the body.

Cancer is a highly heterogeneous disease, with over two hundreddifferent known cancers that affect humans. The causes of cancer arediverse, complex, and only partially understood. Factors that increasethe risk of cancer include tobacco use, dietary factors, certaininfections, exposure to radiation, lack of physical activity, obesity,and environmental pollutants. These factors can directly damage genes orcombine with existing genetic faults within cells to cause cancerousmutations. Approximately 5-10% of cancers can be traced directly toinherited genetic defects.

Cancer is the second leading cause of deaths in the U.S., and the numberof deaths due to cancer continues to grow. Cancer treatment may includesurgery, radiation and/or chemotherapy, based upon the type, locationand dissemination of cancer. Surgery and localized radiation therapy maypresent lower toxicities to healthy cells and tissues, whilechemotherapy is the best treatment option for disseminated cancer,leukemia, lymphoma, and metastasized cancers. Not all tumors respond tochemotherapeutic agents. Other tumors, although initially responsive tochemotherapeutic agents, may develop resistance, with cancer eventuallyrecurring.

Various classes of chemotherapeutic agents have been described. Thesechemotherapeutic agents can be natural products, structurally modifiednatural products, or synthetic chemical or biological agents. Themajority of chemotherapeutic drugs act by interfering with and/orpreventing cell division, or interfering with DNA synthesis or function.Interestingly, novel tyrosine kinase inhibitors, such as imatinibmesylate (Gleevec or Glivec), target a specific molecular abnormality inspecific types of cancer, and their use is thus limited to cancers thatcarry such abnormalities.

Angiogenesis is the physiological process through which new bloodvessels form from pre-existing vessels. This is distinct fromvasculogenesis, which is the de novo formation of endothelial cells frommesoderm cell precursors. Angiogenesis is a normal and vital process ingrowth and development, wound healing and the formation of granulationtissue. However, it is also a fundamental step in the transition oftumors from a benign state to a malignant one. Further, uncontrolledangiogenesis may damage various organs and tissues, such as eyes, skin,heart, blood vessels, lung, gastrointestinal tract and genitourinarytract. Thus, anti-angiogenesis agents (also known as angiogenesisinhibitors) may be used in the treatment of cancer and/or prevention ofuncontrolled angiogenesis.

Cholesterol is a steroidal metabolite that is found in the cellmembranes and transported in the blood plasma of all animals.Cholesterol is an essential structural component of mammalian cellmembranes, where it is required to establish proper membranepermeability and fluidity. In addition, cholesterol is an importantcomponent for the manufacture of bile acids, steroid hormones, andfat-soluble vitamins including vitamins A, D, E and K.

Cholesterol accumulation has been reported in various solid tumors,including prostate cancer and oral cancer. In addition, cholesterolmetabolism is dysregulated in many malignancies, including myeloidleukemia, lung cancer and breast cancer. Specifically, the activity of3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, therate-limiting enzyme in cholesterol biosynthesis, is up-regulated invarious tumors. Malignant cells often have elevated levels ofmevalonate, which formation is catalyzed by HMG-CoA, and consistentlymevalonate treatment was found to promote tumor growth in vivo and tostimulate the proliferation of breast cancer cells. Cholesterolmetabolism is also dysregulated in many hematopoietic malignancies,including acute myeloid leukemia. High cellular cholesterol may in factincrease leukemia cell survival and impart relative resistance totherapy.

Cholesterol derivatives include 7-dehydrocholesterol (7DHC), aprovitamin D present in animals. The presence of 7DHC in skin enableshumans to manufacture vitamin D3 using ultraviolet rays in the sunlight. Increased levels of 7DHC and decreased levels of cholesterol werefound in patients with Smith-Lemli-Opitz syndrome (SLOS), an autosomalrecessive malformation syndrome associated with intellectual disabilityand behavioral problems. Unfortunately, only a few examples of syntheticderivatives of cholesterol are known, and their effects on biosynthetic,metabolic and catabolic pathways of cholesterol remain unexplored.

There is a need in the art to identify novel compounds that can be usedto treat or prevent cancer in a subject. There is also a need in the artto identify novel compounds that can be used to treat or preventuncontrolled angiogenesis in a subject. The present invention addressesand meets these needs.

BRIEF SUMMARY OF THE INVENTION

As described below, the present invention provides novel7-dehydrocholesterol (7DIIC) derivatives that are useful in treating orpreventing cancer, as well as in treating or preventing uncontrolledangiogenesis, in a subject. In certain embodiments of the presentinvention, the subject is a human.

In one aspect, the invention provides compounds, or a salt or solvatethereof.

In another aspect, the invention provides a method of preparingcompounds of the invention, or a salt or solvate thereof.

In yet another aspect, the invention provides a pharmaceuticalcomposition comprising at least one compound of the invention and apharmaceutically acceptable carrier.

In yet another aspect, the invention provides a method of preventing ortreating cancer in a subject in need thereof, the method comprisingadministering to the subject a therapeutically effective amount of atleast one compound of the invention or a salt or solvate thereof.

In yet another aspect, the invention provides a method of preventing,reversing or inhibiting angiogenesis in a subject in need thereof, themethod comprising administering to the subject an effective amount of apharmaceutical composition comprising at least one compound of theinvention or a salt or solvate thereof.

In yet another aspect, the invention provides a method of diagnosing adisease or disorder in a subject in need thereof, the method comprisingadministering to the subject an effective amount of a pharmaceuticalcomposition comprising at least one compound of the invention or a saltor solvate thereof.

In yet another aspect, the invention provides a method of inhibiting theactivity of the vitamin D receptor (also known as calcitriol receptor),the method comprising contacting the receptor with an effective amountof at least one compound of the invention or a salt or solvate thereof.

In yet another aspect, the invention provides a method of inhibiting theactivity of the vitamin D receptor (also known as calcitriol receptor)in a subject in need thereof, the method comprising administering to thesubject an effective amount of a pharmaceutical composition comprisingat least one compound of the invention or a salt or solvate thereof.

In yet another aspect, the invention provides a prepackagedpharmaceutical composition comprising at least one compound of theinvention, or a salt or solvate thereof, an applicator, and aninstructional material for use thereof.

In certain embodiments, the compound is a compound of formula (I), or asalt or solvate thereof:

wherein in (I):

-   -   R₁ is CR₅ or N, wherein:        -   if R₁ is CR₅, then R₃ is selected from the group consisting            of —N(R₅)₂, —NO, —N(R₅)N(R₅)₂, R₆, —N(R₅)—OR₅, —NH—C(═O)R₅,            alkoxy, —OSO₃H, —O(CR₅)_(n)R₆, —O(CR₅)_(n)alkoxy,            —O(CR₅)_(n+1)OII, —OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅,            and —OC(═O)C(R₅)═C(R₅)₂; or R₃ is selected from the group            consisting of ═O and ═S, and H* is omitted; and.        -   if R₁ is N, then R₃ is selected from the group consisting of            N(R₅)₂, —NO, —N(R₅)N(R₅)₂, R₆, —N(R₅)—OR₅, —NH—C(═O)R₅, Cl,            Br, I, alkoxy, mesyl, tosyl, —O(CR₅)_(n)R₆,            —O(CR₅)_(n+1)OR₅, —OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅,            and —OC(═O)C(R₅)═C(R₅)₂;    -   R₂ is selected from the group consisting of O, S, C(R₄)₂, and        N(R₄);    -   each occurrence of R₄ is independently selected from the group        consisting of H, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,        substituted alkynyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, substituted heteroarylalkyl, OR₅, and N(R₅)₂;    -   each occurrence of R₅ is independently selected from the group        consisting of H, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,        substituted alkynyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   R₆ is selected from the group consisting of F, Cl, Br, I, mesyl,        tosyl, —OSi(R₅)₃, —C(═O)OR₅, and —C(═O)R₅;    -   the dotted line is a single or double bond; and,    -   n is an integer ranging from 1 to 10.    -   In certain embodiments, the compound is a compound of formula        (I):

wherein in (I):

-   -   R₁ is CR₅ or N;    -   R₃ is selected from the group consisting of —N(R₅)₂, —NO,        —N(R₅)N(R₅)₂, R₆, —N(R₅)—OR₅, —NH—C(═O)R₅, F, Cl, Br, I,        hydroxy, alkoxy, mesyl, tosyl, —OSO₃H, —O(CR₅)_(n)R₆,        —O(CR₅)_(n)alkoxy, —O(CR₅)_(n+1)OH, —OC(═O)(CR₅)_(n)R₆,        —OC(═O)(CR₅)_(n)OR₅, and —OC(═O)C(R₅)═C(R₅)₂; or R₃ is selected        from the group consisting of ═O and ═S, and H* is omitted;    -   R₂ is selected from the group consisting of O, S, C(R₄)₂, and        N(R₄);    -   each occurrence of R₄ is independently selected from the group        consisting of H, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,        substituted alkynyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, substituted heteroarylalkyl, OR_(S), and        N(R₅)₂;    -   each occurrence of R₅ is independently selected from the group        consisting of H, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,        substituted alkynyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;    -   R₆ is selected from the group consisting of F, Cl, Br, I, mesyl,        tosyl, —OSi(R₅)₃, —C(═O)OR₅, and —C(═O)R₅;    -   the dotted line is a single or double bond; and,    -   n is an integer ranging from 1 to 10.

In certain embodiments, the compound of formula (I) is the compound offormula (Ia), or a salt or solvate thereof:

In certain embodiments, the compound of formula (I) is the compound offormula (Ib), or a salt or solvate thereof:

In certain embodiments, R₁ is N. In other embodiments, R₂ is N(R₄).

In certain embodiments, the compound of formula (I) is the compound offormula (Ic), or a salt or solvate thereof:

In certain embodiments, the compound of formula (I) is the compound offormula (Id), or a salt or solvate thereof:

In certain embodiments, R₃ is selected from the group consisting of—O(CR₅)_(n)R₆, —OC(═O)(CR5)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and—OC(═O)C(R₅)═C(R₅)₂.

In certain embodiments, the compound of formula (I) is selected from thegroup consisting of:

In certain embodiments, R₁ is CR₅. In other embodiments, R₅ is selectedfrom the group consisting of H, alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, arylalkyl, substituted arylalkyl,heteroarylalkyl, and substituted heteroarylalkyl. In yet otherembodiments, R₃ is selected from the group consisting of R₆,—O(CR₅)_(n)R₆, OC(═O)(CR₅)_(n)R₇, and OC(═O)C(R₅)═C(R₅)₂; or R³ isselected from the group consisting of ═O and ═S, and H* is omitted. Inyet other embodiments, n is 1, 2, 3, 4 or 5. In yet other embodiments,the salt is an acid addition salt and is selected from the groupconsisting of sulfate, hydrogen sulfate, hydrochloric, hydrobromic,hydriodic, nitric, carbonic, sulfuric, phosphoric, formic, acetic,propionic, succinic, glycolic, gluconic, lactic, malic, tartaric,citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic,glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic,mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic,benzenesulfonic, pantothenic, trifluoromethanesulfonic,2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic,cyclohexylaminosulfonic, stearic, alginic, -hydroxybutyric, salicylic,galactaric and galacturonic acid, and any combinations thereof. In yetother embodiments, the salt is a base addition salt and is selected fromthe group consisting of calcium, magnesium, potassium, sodium, ammonium,zinc, a basic amine salt, and any combinations thereof, wherein thebasic amine is selected from the group consisting of triethylaminc,diisopropylethylamine, trimethylamine, N,N′-dibenzylethylene-diamine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine,procaine and any combinations thereof.

In certain embodiments, the compound is administered to the subject as apharmaceutical composition comprising at least one compound of theinvention and a pharmaceutically acceptable carrier.

In certain embodiments, the composition further comprises at least oneadditional chemotherapeutic agent selected from the group consisting ofalkylating agents; nitrosoureas; antimetabolites; antitumor antibiotics;plant alkyloids; taxanes; hormonal agents; anti-angiogenesis agents, andmiscellaneous agents. In other embodiments, the composition furthercomprises at least one additional anti-angiogenesis agent selected fromthe group consisting of 2-methoxyestradiol AG3340, angiostatin,antithrombin-III, anti-VEGF antibody, VEGF antagonist, batimastat,bevacizumab, BMS-275291, CA1, canstatin, combretastatin,combretastatin-A4 phosphate, CC-5013, captopril, celecoxib, dalteparin,EMD121974, endostatin, erlotinib, gefitinib, genistein, halofuginone,ID1, ID3, IM862, omatinib mesylate, inducible protein-10,interferon-alpha, interleukin-12, lavendustin-a, LY317615, AE-941,merimastat, mapsin, medroxpregesteron acetate, Meth-1, Meth-2,Neovastat, osteopontin cleaved product, PEX, pigment epithelium growthfactor, platelet growth factor 4, prolactin fragment, proliferin-relatedprotein (PRP), PTK787/ZK222584, recombinant human platelet factor-4,restin, squalamine, SU5416, SU6668, suramin, taxol, tecogalan,thalidomide, thrombospondin, TNP-470, troponin I, vasostatin, VEGF1,VEGF-TRAP and ZD6474. In yet other embodiments, the compound and theagent are co-formulated in the composition.

In certain embodiments, the cancer comprises one selected from the groupconsisting of breast cancer, prostate cancer, ovarian cancer, cervicalcancer, skin cancer, pancreatic cancer, colorectal cancer, renal cancer,liver cancer, brain cancer, lymphoma, leukemia, lung cancer, endometrialcancer, neuroblastoma, and any combinations thereof. In otherembodiments, the cancer comprises one selected from the group consistingof prostate cancer, breast cancer, ovarian cancer, endometrial cancer,medulloblastoma, neuroblastoma, melanoma, and any combinations thereof.

In certain embodiments, the subject is further administered at least oneadditional chemotherapeutic agent. In other embodiments, the subject isfurther administered at least one additional anti-angiogenesis agent. Inyet other embodiments, the compound and the agent are separatelyadministered to the subject. In yet other embodiments, the compound andthe agent are co-administered to the subject. In yet other embodiments,the subject is a mammal. In yet other embodiments, the mammal is ahuman. In yet other embodiments, the composition is administered to thesubject by at least one route selected from the group consisting ofintravenous, oral, inhalational, rectal, vaginal, transdermal,intranasal, buccal, sublingual, parenteral, intrathecal, intragastrical,ophthalmic, pulmonary and topical routes.

In certain embodiments, the method further comprises procuring thecompound of the invention for the subject.

In certain embodiments, the method of preparing a compound of theinvention comprises reacting a compound of formula (II) with a compoundof formula (III):

In other embodiments, the compounds of formula (II) and (III) arereacted in the dark or under reduced light conditions.

In certain embodiments, the instructional material comprisesinstructions for preventing, treating or inhibiting cancer orangiogenesis in a subject. In other embodiments, the prepackagedpharmaceutical composition further comprises at least one additionalchemotherapeutic agent or an antiangiogenesis agent.

Other aspects, embodiments, advantages, and features of the presentinvention will become apparent from the following specification.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description of various embodiments of theinvention will be better understood when read in conjunction with theappended drawings. For the purpose of illustrating the invention, thereare shown in the drawings certain specific embodiments. It should beunderstood, however, that the invention is not limited to the precisearrangements and instrumentalities of the embodiments shown in thedrawings.

FIG. 1 is a bar graph illustrating the cell viability of SKOV-3 cellstreated with 7-dehydrocholesterol and Compound (2).

FIG. 2 is a bar graph illustrating the cytotoxic effect of a compound ofthe invention on SKOV-3 cells.

FIG. 3 is a bar graph illustrating the cytotoxic effect of a compound ofthe invention on ECC-1 cells.

FIG. 4 is a bar graph illustrating the cytotoxic effect of a compound ofthe invention in a panel of ovarian and endometrial cancer,neuroblastoma, breast and prostate cancer cells. Also included are therelative cell viability of a third semester trophoblast treated withCompound (3).

FIG. 5 is a bar graph illustrating the reduction in cytotoxic effects ofa compound of the invention in a panel of ovarian cancer, endometrialcancer, prostate cancer, breast cancer and neuroblastoma uponpretreatment with ascorbic acid suggesting that compounds of theinvention such as compound (3) lead to formation of lethal levels ofreactive oxygen substrates (ROS) or radicals production.

FIGS. 6A and 6B are a set of HPLC profiles of material extracted fromuntreated SKOV-3 cells (FIG. 6A) and SKOV-3 cells treated with Compound(3) (FIG. 6B).

FIG. 7 is a graph illustrating the time-dependent overall tumor size ina xenograph model in nude mice.

FIG. 8 illustrates the antiangiogenic effects of a compound of theinvention via a wound healing assay model based on primary HUVEC cells.

FIG. 9 illustrates the antiangiogenic effects of a compound of theinvention via an ex vivo rat aorta ring assay. MeTC7 inhibited capillaryformation and outgrowth in an ex vivo rat aorta assay (24 hourtreatment).

FIGS. 10A-10F illustrate the finding that compounds of the invention, asexemplified by Compound (3), are selective Vitamin D nuclear receptor(VDR) antagonists, as recited in Example 10. FIG. 10A is a graph thatillustrates antagonistic studies with VDR. FIG. 10B is a graph thatillustrates agonistic studies with VDR. FIG. 10C is a table summarizingselected results of the present invention. FIG. 10D is a graph thatillustrates results for the VDR transactivation assay (as % effect vs.log [Con]). FIGS. 10E-10F are graphs that illustrate the determinationof, respectively, antagonistic and agonistic properties of MeTC7 using aPPAR—coactivator binding assay. FIGS. 11A-11I illustrate the findingthat compounds of the invention, as exemplified by Compound (3), blockgrowth of various cancer cell lines, cause apoptosis and reduce thegrowth of ovarian cancer xenografts in animals, as recited in Example11. FIGS. 11A-11C illustrate viability of selected cell lines before andafter MeTC7 treatment. FIGS. 11D-11E are a set of images illustratingeffects of treatment of cell lines with compounds of the invention. FIG.11F is a graph illustrating tumor size as a function of time (days).FIG. 11G is a graph illustrating survival proportion as a function oftime (days). FIG. 11H is a graph illustrating average weight as afunction of time (days). FIG. 11I is a set of images illustratingeffects of compounds of the invention on xenografts.

FIG. 12 is a set of graphs that illustrate the finding that compounds ofthe invention, as exemplified by Compound (3), reduce the growth ofmedulloblastoma xenografts in animals (Example 12). Exemplificationincludes time-dependent tumor size and body weight.

FIG. 13 is a set of graphs that illustrate the finding that compounds ofthe invention, as exemplified as Compound (3), reduce the growth of BRAFmutant melanoma xenografts in animals, extend survival in these animals(Example 13). Exemplification includes tumor size and survival analysis.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the unexpected identification of novel7-dehydrocholesterol derivatives that are useful in treating orpreventing cancer, as well as in treating or preventing uncontrolledangiogenesis in a subject. In certain embodiments of the presentinvention, the subject is a human.

As demonstrated herein, the compounds of the invention are potentcytotoxic agents against a wide variety of cancel cells. In anon-limiting example, selected compounds of the invention were shown tohave IC₅₀ values ranging from 1 pM to 100 μM against ovarian andendometrial cancer cell lines. Further, in a preliminary cell viabilityassay, a compound of the invention demonstrated potent cytotoxic effectsagainst a panel of cultured endometrial cancer cells (ECC-1, RL-95,AN3CA; IC₅₀<100 nM) and ovarian cancer cells (SKOV-3; IC₅₀<100 nM),prostate cancer cells (PC-3; IC₅₀<100 nM), neuroblastoma cells (SMSKCNR;IC₅₀<10 μM) and breast cancer cells (MCF-7; IC₅₀>20 μM) cells within 24hours. Further, a compound of the invention reduced the cell viabilityof MCF-7 cancer cells (IC₅₀<20 μM) within 72 hours of treatment.

In contrast, a much higher concentration of the compounds of theinvention was required to affect viability of normal trimestertrophoblastic cells (TCL-1), indicating that the compounds of theinvention display high therapeutic and safety.

Without wishing to be limited by any theory, in certain aspects, thecompounds of the invention inhibit lipid synthesis in a cancer cellselectively. Normal cells generally do not perform their own de novolipid synthesis, rather depending on dietary fat uptake. On the otherhand, cancer cells often conduct de novo lipid synthesis to aid theirrapid proliferation and energy needs. In certain embodiments, thecompounds of the invention interfere with the de novo lipid synthesis incancer cells, inhibiting tumor growth and reducing tumor burden, withoutconcomitant off-target toxicity.

Without wishing to be limited by any theory, in certain aspects, thecompounds of the invention act as antagonists of the Vitamin D receptor(also known as calcitriol receptor).

In certain embodiments of the invention, the compounds of the inventionare used to reduce obesity and obesity-induced disorders, such asdiabetes and cardiovascular diseases.

In certain embodiments of the invention, the compounds of the inventionare used to treat cholesterol disorders.

In certain embodiments of the invention, the compounds of the inventionare used to treat patients suffering from Smith-Lemli-Opitz syndrome(SLOS).

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the certain specificmethods and materials are described.

As used herein, each of the following terms has the meaning associatedwith it in this section.

The articles “a” and “an” are used herein to refer to one or to morethan one (i. e., to at least one) of the grammatical object of thearticle. By way of example, “an element” means one element or more thanone element.

“About” as used herein when referring to a measurable value such as anamount, a temporal duration, and the like, is meant to encompassvariations of ±20% or ±10%, more advantageously ±5%, even moreadvantageously ±1%, and still more advantageously ±0.1% from thespecified value, as such variations are appropriate to perform thedisclosed methods.

As used herein, the term “7DHC” refers to 7-dehydrocholesterol.

As used herein, the term “DMSO” refers to dimethylsulfoxide.

As used herein, the term “PBST” refers to phosphate buffered salinesolution with Tween 20.

As used herein, the term “DMEM” refers to Dulbecco's modified Eagle'smedium.

As used herein, the term “RPMI” refers to Roswell Park MemorialInstitute medium.

As used herein, the term “MTS” refers to(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)or a salt thereof.

A “disease” as used herein is a state of health of an animal wherein theanimal cannot maintain homeostasis, and wherein if the disease is notameliorated then the animal's health continues to deteriorate.

A “disorder” as used herein in an animal is a state of health in whichthe animal is able to maintain homeostasis, but in which the animal'sstate of health is less favorable than it would be in the absence of thedisorder. Left untreated, a disorder does not necessarily cause afurther decrease in the animal's state of health.

A disease or disorder is “alleviated” if the severity of a symptom ofthe disease or disorder, the frequency with which such a symptom isexperienced by a patient, or both, is reduced.

As used herein, the term “cancer” is defined as disease characterized bythe rapid and uncontrolled growth of aberrant cells. Cancer cells canspread locally or through the bloodstream and lymphatic system to otherparts of the body. Examples of various cancers include, but are notlimited to, breast cancer, prostate cancer, ovarian cancer, cervicalcancer, skin cancer, pancreatic cancer, colorectal cancer, renal cancer,liver cancer, brain cancer, lymphoma, leukemia, lung cancer, endometrialcancer, neuroblastoma, medulloblastoma, melanoma, and the like.

Non-limiting examples of solid cancers are breast, ovarian, endometrial,cervical, neuroblastoma, medulloblastoma, lung cancer, colon cancer, CNScancer, melanoma, renal, prostate, medulloblastoma, head and neckcancer, esophagus cancer, pancreatic cancer, skin cancer, thyroidalcancer, peripheral nerve sheath cancer, ependymoma, cranaiopharyngioma,astrocytoma (juvenile pilocytic astrocytoma, subependymal giant cellastrocytoma, pleimorphic xanthoastrocytoma, analplastic astrocytoma, orgliomatosis cerebri), meningioma, germinoma, glioma, mixed glioma,choroid plexus tumor, oligodendroglioma, peripheral neuroectodermaltumors, CNS lymphoma, pituitary adenoma, or shwannoma.

As used herein, the term “carcinoma” refers to any cancer of epithelialorigin. Non-limiting examples of carcinomas include, but are not limitedto, acinar carcinoma, acinous carcinoma, alveolar adenocarcinoma,carcinoma adenomatosum, adenocarcinoma, carcinoma of adrenal cortex,alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma,carcinoma basocellulare, basaloid carcinoma, basosquamous cellcarcinoma, breast carcinoma, branchioalveolar carcinoma, bronchiolarcarcinoma, cerebrifoim carcinoma, cholangiocellular carcinoma, chorioniccarcinoma, colloid carcinoma, comedo carcinoma, corpus carcinoma,cribriform carcinoma, carcinoma en cuirasse, carcinoma cutaneum,cylindrical carcinoma, cylindrical cell carcinoma, duct carcinoma,carcinoma durum, embryonal carcinoma, encephaloid carcinoma, epibulbarcarcinoma, epidermoid carcinoma, carcinoma epitheliate adenoids,carcinoma exulcere, carcinoma fibrosum, gelatinform carcinoma,gelatinous carcinoma, giant cell carcinoma, gigantocellulare, glandularcarcinoma, granulose cell carcinoma, hair matrix carcinoma, hematoidcarcinoma, hepatocellular carcinoma, Hurthle cell carcinoma, hyalinecarcinoma, hypernephroid carcinoma, infantile embryonal carcinoma,carcinoma in situ, intraepidermal carcinoma, intraepithelial carcinoma,Krompecher's carcinoma, Kulchitzky-cell carcinoma, lentivular carcinoma,carcinoma lenticulare, lipomatous carcinoma, lymphoepithelial carcinoma,carcinoma mastotoids, carcinoma medullare, medullary carcinoma,carcinoma melanodes, melanotonic carcinoma, mucinous carcinoma,carcinoma muciparum, carcinoma mucocullare, mucoepidermoid carcinoma,mucous carcinoma, carcinoma myxomatodes, masopharyngeal carcinoma,carcinoma nigrum, oat cell carcinoma, carcinoma ossificans, osteroidcarcinoma, ovarian carcinoma, papillary carcinoma, periportal carcinoma,preinvasive carcinoma, prostate carcinoma, renal cell carcinoma ofkidney, reserve cell carcinoma, carcinoma sarcomatodes, scheinderiancarcinoma, scirrhous carcinoma, carcinoma scrota, signet-ring cellcarcinoma, carcinoma simplex, small cell carcinoma, solandoid carcinoma,spheroidal cell carcinoma, spindle cell carcinoma, carcinoma spongiosum,squamous carcinoma, squamous cell carcinoma, string carcinoma, carcinomatelangiectaticum, carcinoma telangiectodes, transitional cell carcinoma,carcinoma tuberrosum, tuberous carcinoma, verrucous carcinoma, orcarcinoma vilosum.

As used herein, the term “sarcoma” refers to any mesenchymal neoplasmthat arises in bone and soft tissues. Non-limiting examples of sarcomasinclude liposarcomas (including myxoid liposarcomas and pleiomorphicliposarcomas), leiomyosarcomas, rhabdomyosarcomas, neurofibrosarcomas,malignant peripheral nerve sheath tumors, Edwing's tumors (includingEdwing's sarcoma of bone, extraskeletal or non-bone) and primitiveneuroectodermal tumors (PNET), synovial sarcoma, hemangiendothelioma,fibrosarcoma, desmoids tumors, dermatofibrosarcoma protuberance (DFSP),malignant fibrous histiocytoma(MFH), hemangiopericytoma, malignantmesenchymoma, alveolar soft-part sarcoma, epitheloid sarcoma, clear cellsarcoma, desmoplastic small cell tumor, gastrointestinal stromal tumor(GIST) and osteosarcoma, and chondrosarcoma.

As used herein, the term “refractory cancer” refers to a cancer that isresistant to the ordinary standards of care prescribed. These cancers,although potentially initially responsive to treatment, recur and/or maybe completely non-responsive to the treatment.

As used herein, the term “immunogenic cancer” refers to cancers selectedfrom the group consisting of malignant melanoma and renal cellcarcinoma, Mantel cell lymphoma, follicular lymphoma, diffuse largeB-cell lymphoma, T-cell acute lymphoblastic leukemia, Burkitt Lymphoma,myeloma, immunocytoma, acute promyelocytic leukemia, chronicmyeloid/acute lyphoblastic leukemia, acute leukemia, B-cell acutelymphoblastic leukemia, anaplastic large cell leukemia, myelodysplasticsyndrome/acute myeloid leukemia, non-Hodgkin's lymphoma, chroniclymphocytic leukemia, acute myelogenous leukemia (AML), common (pre-B)acute lymphocytic leukemia, malignant melanoma, T-cell lymphoma,leukemia, B-cell lymphoma, epithelial malignancies, lymphoidmalignancies, gynecologic carcinoma, biliary adenocarcinomas and ductaladenocarcinomas of the pancreas.

As used herein, the term “chemotherapeutic agent” refers to a compoundor composition that may be used to treat or prevent cancer. Non-limitingexamples of these agents are DNA damaging agents, such astopoisomeraseinhibitors (for example, etoposide, camptothccin, topotecan,irrinotecan, teniposide, mitoxantrone), anti-microtubule agents (forexample, vincristine, vinblastine), antimetabolite agents (for example,cytarabine, methotrexate, hydroxyurea, 5-fluorouracil, flouridine,6-thioguanine, 6-mercaptompurine, fludaribine, pentostatin,cholorodeoxyadenosine), DNA alkylating agents (for example, cisplatin,mecholorethamine, cyclophosphamide, ifosphamide, melphalan,chlorumbucil, busulfan, thiotepa, carmustine, lomustine, carboplatin,dacarbazine, procarbazine) and DNA strand break inducing agents (forexample, bleomycin, doxarubicine, daunorubicine, idarubicine, mitomycinC). Chemotherapeutic agents include but are not limited to avicin,aclarubicin, acodazole, acronine, adozelesin, adriamycin, aldesleukin,alitretnoin, allopurinl sodium, altretamine, ambomycin, amitantroneacetate, aminoglutethimide, amscrine, anastrazole, annoceousacetogenins, anthramycin, asimicin, asparaginase, asperlin, azacitidine,azetepa, azotomycin, batimstat, benzodepa, bexarotenc, bicalutamide,bisantrene, bisanafide, bizelesin, bleomycin, brequinar, brompirimine,bullatacin, busulfan, cabergoline, cactinomycin, calusterone,caracemide, carbetimer, carbopltin, carmustine, carubicin, carzelesin,cedefingol, chlorumbucil, celecoxib, cirolemycin, cisplatin,cladiribine, crisnatol, cyclophosphamide, cytarabine, dacarbazine, DACA,dactinomycin, daunorubicine, daunomycin, decitabine, denileukine,dexormaplatin, dezaguanine, diaziqone, docetaxel, doxarubicin,droloxifene, dromostalone, duazomycin, edatrexate, eflornithin,elsamitrucin, estramustine, etanidazole, etoposide, etropine, fadrozole,fazarabine, feneretinide, floxuridine, fludarabine, flurouracil,fluorocitabine, 5-FdUMP, fosquidone, fosteuecine, FK-317, FK-973,FR-66979, FR-900482, gemcitabine, gemtuzumab, ozogamicin, Gold Au198,goserelin, guanacone, hydroxyurea, idarubicine, ilmofosine, interferonalpha and analogs, iprolatin, irinotecan, lanreotide, letrozole,leuprolide, liarozole, lometrexol, lomustine, losoxantrone, masoprocol,maytansine, maturedepa, mecholoroethamine, megesterol, melengesterol,melphalan, menogaril, metoprine, mycophenolic acid, mitindomide,mitocarcin, mitogillin, mitomalacin, mitomycin, mitomycin C, mitosper,mitotane, mitoxantrone, nocodazole, nogalamycin, oprelvekin, ormaplatin,profiromycin, oxisuran, paclitaxel, pamidronate, pegaspargase,pcliomycin, pentamustin, peplomycin, perfosfamide, pipobroman,piposulfan, piroxantrone, plicamycin, plomestane, porfimer,prednimustin, procarbazine, puromycin, pyrazofurin, riboprine,rogletimide, rituximab, rolliniastatin, safingol, samarium, semustine,simtrazene, sparfosate, sparcomycin, sulphofenur, spirogermanium,spiromustin, spiroplatin, squamocin, squamotacin, streptozocin,streptonigrin, SrCl₂, talosmycin, taxane, taxoid, tecoglan, temoprofin,tegafur, teloxantrone, teniposide, terxirone, testolactone, thiamiprine,thiotepa, thymitaq, tomudex, tiazofurin, tirapamazinc, Top-53,topotecan, toremixifine, trastuzumab, trestolone, tricribine,trimetrexate, tricribine, trimetrexate glucuronate, triptorelin,tubulozole, uracil mustard, valrubicine, uredepa, vapreotide,vinblastin, vincristine, vindesin, vinepidine, zinostatin, vinglycinate,vinleurosine, vinorelbine, vinrosidine, vinzolidine, vorozole,zeniplatin, zorubicin, 2-chlorodeoxyrubicine, 2′-deoxyformycin, CEP-751,raltitrexed, N-propargyl-5,8-didezafolic acid,2-chloro-2′-arabinofluoro-2′-deoxyadenosine, 2-chloro-2′-deoxyadenosine,9- aminocamptothecin anisomycin, trichostatin, hPRL-G129R, linomide,sulfur mustard, N-methyl-N-nitrosourea, fotemustine, streptozotocin,bisplatinum, temozolomide, mitozolomide, AZQ, ormaplatin, CI-973,DWA2114R, JM216, JM335, tomudex, azacitidine, cytrabincine, gemcitabine,6-mercaptopurine, teniposide, hypoxanthine, doxorubicine, CPT-11,daunorubicine, darubicin, epirubicine, nitrogen mustard, losoxantronc,dicarbazine, amscrinc, pyrazoloacridinc, all trans retinol,14-hydroxy-retro-retinol, all-trans retinoic acid, N-(4-hydroxyphenyl)rertinamide, 13-cisretinoic acid, 3-methyl TTNEB, 9-cisretenoic acid,fludarabine, and 2-Cda.

Other chemotherapeutic agent include: adecylpenol,20-epi-1,25-dihydroxyvitamin-D3, 5-ethynyl uracil, abiraterone,aclarubicine, acylfulvene, adozelecin, aldesleukin, antagonists,altretamine, ambumastine, amidox, amifostine, amino levulinic acid,anagralide, anastrozole, andrographolide, antagonist D, antarelix,anti-dorsalizing morphogenetic protein-1, antiandrogen, antiestrogen,antineoplastone, antisense oligonucleotides, aphidicolin, apoptosis genemodulators, apotosis regulators, apurinic acid, ara-cdp-dl-PTBA,arginine aminase, asulacrine, atamestine, atrimustine, axinamastine 1and axinamastine 2, axinamastine 3, azasetron, azatoxin, azatyrosine,baccatin III derivatives, balanol, BCR/ABL antagonist, benzochlorins,benzoylsaurosporine, beta lactam derivatives, beta-alethine, perillylalcohol, phenozenomycin, phenyl acetate, phosphatase inhibitors,picibanil, pilocarbine and salts or analogs thereof, pirarubucin,piritrexim, placetin A, placetin B, plasminogen activator inhibitor,platinum complex, phenyl ethyl isothiocyanate and analogs thereof,platinum compounds, platinum triamine complex, podophylotoxin, porfimersodium, porphyromycin, propyl bis acridones, prostaglnadins J2, proteaseinhibitors, protein A based immune modulators, PKC ihibitors,microalgal, protein tyrosine phosphatase inhibitors, purine neucleosidephosphorylase inhibitors, purpurins, pyrazoloacridines, pyridoxylatedhaemoglobin polyoxyethylene conjugate, raf antagonists, raltitrexed,ramosetron, ras farnesyl protein tranaferase inhibitors, ras inhibitors,ras-GAP inhibitors, ratellitptine demethylated, Rhenium Re186etidronate, rhizoxine, ribozyme, RH retinide, rogletimide,rosagliatazone and analogs and derivatives thereof, rohitukine,romurtide, mquinimex, rubiginone B1, ruboxyl, safingol, saintopin,SarCNU, sarcophytol A, sargrmostim, sdi 1 mimetics, semustine,senescence derived inhibitor 1, sense oligonucleotide, signaltransduction inhibitors, signal transduction modulators, single chainantigen binfing protein, sizofiran, sobuzoxane, sodium borocaptate,sodium phenyl acetate, solverol, somatomedin binding protein, sonermin,sparfosic acid, spicamycin D, spiromustin, splenopentine, spongistatin1, squalamine, stem cell inhibitor, stem cell division inhibitor,stipiamide, stromelysin, sulfinosine, superactive vasoactive intestinalpeptide antagonists, suradista, siramin, swainsonine, syntheticglycosaminoglycans, tallimustine, tamoxifen methiodide, tauromustine,tazarotene, tacogalan sodium, tegafur, tellurapyrilium, telomeraseinhibitors, temoporfin, tmeozolomide, teniposide, tetrachlorodecaoxide,tetrazomine, thaliblastine, thalidomide, thiocoraline, thrombopoetin andmimetics thereof, thymalfasin, thymopoetin receptor agonist,thymotrinan, thyroid stimulating harmonc, tin ethyl etiopurpin,tirapazamine, titanocene and salts thereof, topotecan, topsentin,toremifene, totipotent stem cell factors, translation inhibitors,tretinoin, triacetyluridine, tricribine, trimetrexate, triptorelin,tropisetron, turosteride, tyrosine kinase inhibitors, tyrphostins, UBCinhibitors, ubenimex, urogenital sinus derived growth inhibitory factor,urokinase receptor antagonists, vapreotide, variolin vector system,erythrocyte gene therapy, velaresol, veramine, verdins, verteporfin,vinorelbine, vinxaltine, vitaxin, vorozol, zanoterone, zeniplatin,zilascorb and zinostatin.

Other chemotherapeutic agents include antiproliferative agents (e.g.,piritrexim isothiocyanate), antiprostatic hypertrophy agents(sitogluside), Benign prostatic hyperplasia therapy agents (e.g.,tomsulosine, RBX2258), prostate growth inhibitory agents (pentomone) andradioactive agents: fibrinogen I125, fludeoxyglucose F18, flurodopa F18,insulin I125, iobenguane I123, iodipamide sodium I131, iodoantipyrineI131, iodocholesterol I131, iodopyracet I125, iofetamine HCL I123,iomethin I131, iomethin I131, iothalamate sodium I125, iothalamate I131,iotyrosine I131, liothyronine I125, merosproprol Hg197, methylioodobenzo guanine (MIBG-I131 or MIBG-I123) selenomethionine Se75,technetium Tc99m furifosmin, technetium Tc99m gluceptate, Tc99mbiscisate, Tc99m disofenin, TC99m gluceptate, Tc99m lidofenin, Tc99mmebrofenin, Tc99m medronate and sodium salts thereof, Tc99m mertiatide,Tc99m oxidronate, Tc99m pentetate and salts thereof, Tc99m sestambi,Tc99m siboroxime, Tc99m succimer, Tc99m sulfur colloid, Tc 99mteboroxime, Tc 99m tetrofosmin, Tc99m tiatide, thyroxine I125, thyroxineI131, tolpovidone I131, triolein I125, treoline I125, and treoline 131.

Another category of chemotherapeutic agents is anticancer supplementarypotentiating agents, e.g., antidepressant drugs (imipramine,desipramine, amitryptyline, clomipramine, trimipramine, doxepin,nortryptylinc, protryptylinc, amoxapine, and maprotiline), orno-trycyclic anti-depressant drugs (sertaline, trazodone andcitalopram), Ca⁺⁺ antagonists (vermapil, nifedipine, nitrendipine andcaroverine), calmodulin inhibitors (prenylamine, trifluroperazine andclomipramine), amphotericin B, triparanol analogs (e.g., tomoxifene),antiarrythmic drugs (e.g., quinidine), antihypertensive drugs (e.g.,resepine), thiol depleters (e.g., buthionine and sulofoximine) andmultiple drug resistance reducing agents such as cremaphor EL.

In certain embodiments, chemotherapeutic agents include annoceousacetogenins, ascimicin, rolliniastatin, guanocone, squamocin,bullatacin, squamotacin, axanes, baccatin, and taxanes (Paclitaxel anddocetaxel).

In certain embodiments, chemotherapeutic agents include anti-CD20 mAB,rituximab, rituxan, tositumoman, Bexxar, anti-HER2, trastuzumab,Herceptin, MDX20, antiCA125 mAB, antiHE4 mAB, oregovomab mAB, B43.13mAB, Ovarex, Breva-REX, AR54, GivaRex, ProstaRex mAB, MDX447, gemtuzumabozoggamycin, Mylotarg, CMA-676, anti-CD33 mAB, anti-tissue factorprotein, Sunol, IOR-C5, C5, anti-EGFR mAB, anti-IFR1R mAR, MDX-447,anti-17-1A mAR, edrecolomab mAR, Panorex, anti-CD20 mAR (Y-90 lebelled),ibritumomab tiuxetan (IDEC-Y2B8), zevalin, and anti-idiotypic mAB.

As used herein, the term “uncontrolled angiogenesis” refers toangiogenesis that is not part of the normal or healthy development ofnovel blood vessels in a subject. Uncontrolled angiogenesis may beassociated with cancer, ocular disease (for example, maculardegeneration, maculopathy, diabetic retinopathy or retinopathy ofprematurity (retrolental fibroplasia)), skin disease (for example,infantile hemangioma, verruca vulgaris, psoriasis, neurofibromatosis, orepidermolysis bullosa), autoimmune disease (for example, rheumatoidarthritis), gynecologic disease (for example, endometrial polyp,endometriosis, dysfunctional uterine bleeding, ovarian hyperstimulationsyndrome, polycystic ovarian syndrome (PCO), or preeclamsia),cardiovascular disease (for example, coronary artery disease, ischemiccardiomyopathy, myocardial Ischemia, arteriosclerosis, atherosclerosis,athelosclerotic plaque, neovascularization, arterial occlusive disease,ischemia, ischemic ulcers, ischemic or post myocardial ischemiarevascularization, peripheral vascular disease or intermittentclaudication), gastrointestinal disease (for example, Crohn's diseaseand ulcerative colitis), buerger disease, thromboangitis obliterans,arteosclerosis obliterans, ischemic ulcers, multiple sclerosis,idiopathic pulmonary fibrosis. HIV infection, plantar faciitis,Von-Hippel Landou disease, CNS hemangioblastoma, retinalhemangioblastoma, thyroiditis, benign prostate hyperplasia,glomerulonephritis, ectopic pregnancy, and ectopic bone formation orkeloid. In certain embodiments, the cancer may be biliary tract cancer,bladder cancer, bone cancer, brain cancer, choriocarcinoma, breastcancer, cervical cancer, colon and rectum cancer, connective tissuecancer, cancer of digestive system, endometrial, esophageal, eye cancer,fibromael, cancer of head and neck, gastric cancer, intra-epithelialneoplasm, kidney cancer, larynx cancer, leukemia including acute myeloidleukemia, acute lymphoid leukemia, chronic lymphoid leukemia, livercancer, lung cancer (for example, small cell and non-small cell),lymphoma including Hodgkins or non-Hodgkins), melanoma, oral cavitycancer (lip, tongue, mouth and pharynx), ovarian cancer, pancreaticcancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, rectalcancer, renal cancer and cancers of the respiratory tract, sarcoma, skincancer, stomach cancer, testicular cancer, thyroid cancer, uterinecancer, cancers of the urinary system, a sarcoma or carcinoma.

The terms “patient,” “subject” or “individual” are used interchangeablyherein, and refer to any animal, or cells thereof whether in vitro or insitu, amenable to the methods described herein. In a non-limitingembodiment, the patient, subject or individual is a human.

As used herein, the term “procure” or “procuring” as relating to asubject in need of being administered a therapeutically active compoundrefers to the act of synthesizing, packaging, prescribing, purchasingand/or providing the compound so that the subject may be administeredthe compound.

As used herein, the term “composition” or “pharmaceutical composition”refers to a mixture of at least one compound useful within the inventionwith a pharmaceutically acceptable carrier. The pharmaceuticalcomposition facilitates administration of the compound to a patient orsubject. Multiple techniques of administering a compound exist in theart including, but not limited to, intravenous, oral, aerosol,parenteral, ophthalmic, pulmonary and topical administration.

A “therapeutic” treatment is a treatment administered to a subject whoexhibits signs of pathology, for the purpose of diminishing oreliminating those signs.

As used herein, the term “treatment” or “treating” is defined as theapplication or administration of a therapeutic agent, i.e., a compoundof the invention (alone or in combination with another pharmaceuticalagent), to a patient, or application or administration of a therapeuticagent to an isolated tissue or cell line from a patient (e.g., fordiagnosis or ex vivo applications), who has a condition contemplatedherein, a symptom of a condition contemplated herein or the potential todevelop a condition contemplated herein, with the purpose to cure, heal,alleviate, relieve, alter, remedy, ameliorate, improve or affect acondition contemplated herein, the symptoms of a condition contemplatedherein or the potential to develop a condition contemplated herein. Suchtreatments may be specifically tailored or modified, based on knowledgeobtained from the field of pharmacogenomics.

The term “prevent,” “preventing” or “prevention,” as used herein, meansavoiding or delaying the onset of symptoms associated with a disease orcondition in a subject that has not developed such symptoms at the timethe administering of an agent or compound commences.

As used herein, the terms “effective amount,” “pharmaceuticallyeffective amount” and “therapeutically effective amount” refer to anontoxic but sufficient amount of an agent to provide the desiredbiological result. That result may be reduction and/or alleviation ofthe signs, symptoms, or causes of a disease, or any other desiredalteration of a biological system. An appropriate therapeutic amount inany individual case may be determined by one of ordinary skill in theart using routine experimentation.

As used herein, the term “pharmaceutically acceptable” refers to amaterial, such as a carrier or diluent, which does not abrogate thebiological activity or properties of the compound, and is relativelynon-toxic, i.e., the material may be administered to an individualwithout causing undesirable biological effects or interacting in adeleterious manner with any of the components of the composition inwhich it is contained.

As used herein, the language “pharmaceutically acceptable salt” refersto a salt of the administered compounds prepared from pharmaceuticallyacceptable non-toxic acids, including inorganic acids, organic acids,solvates, hydrates, or calthrates thereof.

As used herein, the term “pharmaceutically acceptable carrier” means apharmaceutically acceptable material, composition or carrier, such as aliquid or solid filler, stabilizer, dispersing agent, suspending agent,diluent, excipient, thickening agent, solvent or encapsulating material,involved in carrying or transporting a compound useful within theinvention within or to the patient such that it may perform its intendedfunction. Typically, such constructs are carried or transported from oneorgan, or portion of the body, to another organ, or portion of the body.Each carrier is “acceptable” in the sense of being compatible with theother ingredients of the formulation, including the compound usefulwithin the invention, and not injurious to the patient. Some examples ofmaterials that may serve as pharmaceutically acceptable carriersinclude: sugars, such as lactose, glucose and sucrose; starches, such ascorn starch and potato starch; cellulose, and its derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients, such as cocoabutter and suppository waxes; oils, such as peanut oil, cottonseed oil,safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols,such as propylene glycol; polyols, such as glycerin, sorbitol, mannitoland polyethylene glycol; esters, such as ethyl oleate and ethyl laurate;agar; buffering agents, such as magnesium hydroxide and aluminumhydroxide; surface active agents; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffersolutions; and other non-toxic compatible substances employed inpharmaceutical formulations. As used herein, “pharmaceuticallyacceptable carrier” also includes any and all coatings, antibacterialand antifungal agents, and absorption delaying agents, and the like thatare compatible with the activity of the compound useful within theinvention, and are physiologically acceptable to the patient.Supplementary active compounds may also be incorporated into thecompositions. The “pharmaceutically acceptable carrier” may furtherinclude a pharmaceutically acceptable salt of the compound useful withinthe invention. Other additional ingredients that may be included in thepharmaceutical compositions used in the practice of the invention areknown in the art and described, for example in Remington'sPharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985. Easton,Pa.), which is incorporated herein by reference.

In one aspect, the terms “co-administered” and “co-administration” asrelating to a subject refer to administering to the subject a compoundof the invention or salt thereof along with a compound that may alsotreat the disorders or diseases contemplated within the invention. Inone embodiment, the co-administered compounds are administeredseparately, or in any kind of combination as part of a singletherapeutic approach. The co-administered compound may be formulated inany kind of combinations as mixtures of solids and liquids under avariety of solid, gel, and liquid formulations, and as a solution.

By the term “specifically bind” or “specifically binds,” as used herein,is meant that a first molecule preferentially binds to a second molecule(e.g., a particular receptor or enzyme), but does not necessarily bindonly to that second molecule.

As used herein, the term “VDR” refers to Vitamin D nuclear receptor.

As used herein, the term “alkyl,” by itself or as part of anothersubstituent means, unless otherwise stated, a straight or branched chainhydrocarbon having the number of carbon atoms designated (i.e., C₁-C₁₀means one to ten carbon atoms) and includes straight, branched chain, orcyclic substituent groups. Examples include methyl, ethyl, propyl,isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, andcyclopropylmethyl. Certain specific examples include (C₁-C₆)alkyl, suchas, but not limited to, ethyl, methyl, isopropyl, isobutyl, n-pentyl,n-hexyl and cyclopropylmethyl.

As used herein, the term “cycloalkyl,” by itself or as part of anothersubstituent means, unless otherwise stated, a cyclic chain hydrocarbonhaving the number of carbon atoms designated (i.e., C₃-C₆ means a cyclicgroup comprising a ring group consisting of three to six carbon atoms)and includes straight, branched chain or cyclic substituent groups.Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and cyclooctyl. Certain specific examples include(C₃-C₆)cycloalkyl, such as, but not limited to, cyclopropyl, cyclobutyl,cyclopentyl and cyclohexyl.

As used herein, the term “alkenyl,” employed alone or in combinationwith other terms, means, unless otherwise stated, a stablemono-unsaturated or di-unsaturated straight chain or branched chainhydrocarbon group having the stated number of carbon atoms. Examplesinclude vinyl, propenyl (or allyl), crotyl, isopentenyl, butadienyl,1,3-pentadienyl, 1,4-pentadienyl, and the higher homologs and isomers. Afunctional group representing an alkene is exemplified by —CH₂—CH═CH₂.

As used herein, the term “alkynyl,” employed alone or in combinationwith other terms, means, unless otherwise stated, a stable straightchain or branched chain hydrocarbon group with a triple carbon-carbonbond, having the stated number of carbon atoms. Non-limiting examplesinclude ethynyl and propynyl, and the higher homologs and isomers. Theterm “propargylic” refers to a group exemplified by —CH₂—C CH. The term“homopropargylic” refers to a group exemplified by —CH₂CH₂—C CH. Theterm “substituted propargylic” refers to a group exemplified by —CR₂—CCR, wherein each occurrence of R is independently H, alkyl, substitutedalkyl, alkenyl or substituted alkenyl, with the proviso that at leastone R group is not hydrogen. The teen “substituted homopropargylic”refers to a group exemplified by —CR₂CR₂—C CR, wherein each occurrenceof R is independently II, alkyl, substituted alkyl, alkenyl orsubstituted alkenyl, with the proviso that at least one R group is nothydrogen.

As used herein, the term “substituted alkyl,” “substituted cycloalkyl,”“substituted alkenyl” or “substituted alkynyl” means alkyl, cycloalkyl,alkenyl or alkynyl, as defined above, substituted by one, two or threesubstituents selected from the group consisting of halogen, —OH, alkoxy,tetrahydro-2-H-pyranyl, —NH₂, —N(CH₃)₂, (1-methyl-imidazol-2-yl),pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, —C(═O)OH, trifluoromethyl, —CN, —C(═O)O(C₁-C₄)alkyl, —C(═O)NH₂, —C(═O)NH(C₁-C₄)alkyl,—C(═O)N((C₁-C₄)alkyl)₂, —SO₂NH₂, —C(═NH)NH₂, and —NO₂, advantageouslycontaining one or two substituents selected from halogen, —OH, alkoxy,—NH₂, trifluoromethyl, —N(CH₃)₂, and —C(═O)OH, more advantageouslyselected from halogen, alkoxy and —OH. Examples of substituted alkylsinclude, but are not limited to, 2,2-difluoropropyl,2-carboxycyclopentyl and 3-chloropropyl.

As used herein, the term “alkoxy” employed alone or in combination withother terms means, unless otherwise stated, an alkyl group having thedesignated number of carbon atoms, as defined above, connected to therest of the molecule via an oxygen atom, such as, for example, methoxy,ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and the higher homologs andisomers. In certain embodiments, alkoxy includes (C₁-C₃)alkoxy, such as,but not limited to, ethoxy and methoxy.

As used herein, the term “halo” or “halogen” alone or as part of anothersubstituent means, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom, advantageously, fluorine, chlorine, or bromine,more advantageously, fluorine or chlorine.

As used herein, the term “heteroalkyl” by itself or in combination withanother term means, unless otherwise stated, a stable straight orbranched chain alkyl group consisting of the stated number of carbonatoms and one or two heteroatoms selected from the group consisting ofO, N, and S, and wherein the nitrogen and sulfur atoms may be optionallyoxidized and the nitrogen heteroatom may be optionally quaternized. Theheteroatom(s) may be placed at any position of the heteroalkyl group,including between the rest of the heteroalkyl group and the fragment towhich it is attached, as well as attached to the most distal carbon atomin the heteroalkyl group. Examples include: —O—CH₂—CH₂—CH₃,—CH₂—CH₂—CH₂—OH, —CH₂—CH₂—NH-CH₃, —CH₂—S—CH₂—CH₃, and —CH₂CH₂—S(═O)—CH₃.Up to two heteroatoms may be consecutive, such as, for example,—CH₂—NH—OCH₃, or —CH₂—CH₂—S—S—CH₃.

As used herein, the term “heteroalkenyl” by itself or in combinationwith another term means, unless otherwise stated, a stable straight orbranched chain monounsaturated or di-unsaturated hydrocarbon groupconsisting of the stated number of carbon atoms and one or twoheteroatoms selected from the group consisting of O, N, and S, andwherein the nitrogen and sulfur atoms may optionally be oxidized and thenitrogen heteroatom may optionally be quaternized. Up to two heteroatomsmay be placed consecutively. Examples include —CH═CH—O—CH₃,—CH═CH—CH₂—OH, —CH₂—CH═N—OCH₃, —CH═CH—N(CH₃)—CH₃, and —CH₂—CH═CH—CH₂—SH.

As used herein, the term “aromatic” refers to a carbocycle orheterocycle with one or more polyunsaturated rings and having aromaticcharacter, i.e. having (4n+2) delocalized π (pi) electrons, where n isan integer.

As used herein, the term “aryl,” employed alone or in combination withother terms, means, unless otherwise stated, a carbocyclic aromaticsystem containing one or more rings (typically one, two or three rings)wherein such rings may be attached together in a pendent manner, such asa biphenyl, or may be fused, such as naphthalene. Examples includephenyl, anthracyl, and naphthyl. In certain embodiments, aryl includesphenyl and naphthyl, in particular, phenyl.

As used herein, the term “aryl-(C₁-C₃)alkyl” means a functional groupwherein a one to three carbon alkylene chain is attached to an arylgroup, e.g., —CH₂CH₂-phenyl or —CH₂-phenyl (benzyl). Examples includedaryl-CH₂— and aryl-CH(CH₃)—. The term “substituted aryl-(C₁-C₃)alkyl”means an aryl-(C₁-C₃)alkyl functional group in which the aryl group issubstituted. Specific examples include substituted aryl(CH₂)—.Similarly, the term “heteroaryl-(C₁-C₃)alkyl” means a functional groupwherein a one to three carbon alkylene chain is attached to a heteroarylgroup, e.g., —CH₂CH₂-pyridyl. One embodiment is heteroaryl-(CH₂)—. Theterm “substituted heteroaryl-(C₁-C₃)alkyl” means aheteroaryl-(C₁-C₃)alkyl functional group in which the heteroaryl groupis substituted. Specific examples include substitutedheteroaryl-(CII₂)-.

As used herein, the term “heterocycle” or “heterocyclyl” or“heterocyclic” by itself or as part of another substituent means, unlessotherwise stated, an unsubstituted or substituted, stable, mono- ormulti-cyclic heterocyclic ring system that consists of carbon atoms andat least one heteroatom selected from the group consisting of N, O, andS, and wherein the nitrogen and sulfur heteroatoms may be optionallyoxidized, and the nitrogen atom may be optionally quaternized. Theheterocyclic system may be attached, unless otherwise stated, at anyheteroatom or carbon atom that affords a stable structure. A heterocyclemay be aromatic or non-aromatic in nature. In one embodiment, theheterocycle is a heteroaryl.

As used herein, the term “heteroaryl” or “heteroaromatic” refers to aheterocycle having aromatic character. A polycyclic heteroaryl mayinclude one or more rings that are partially saturated. Examples includetetrahydroquinoline and 2,3-dihydrobenzofuryl.

Examples of non-aromatic heterocycles include monocyclic groups such asaziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine,pyrroline, imidazoline, pyrazolidine, dioxolane, sulfolane,2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran, thiophane,piperidine, 1,2,3,6-tetrahydropyridine, 1,4-dihydropyridine, piperazine,morpholine, thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran,1,4-dioxane, 1,3-dioxane, homopiperazine, homopiperidine, 1,3-dioxepane,4,7-dihydro-1,3-dioxepin and hexamethyleneoxide.

Examples of heteroaryl groups include pyridyl, pyrazinyl, pyrimidinyl(such as, but not limited to, 2- and 4-pyrimidinyl), pyridazinyl,thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl,isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl,tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,3,4-thiadiazolyland 1,3,4-oxadiazolyl.

Examples of polycyclic heterocycles include indolyl (such as, but notlimited to, 3-, 4-, 5-, 6- and 7-indolyl), indolinyl, quinolyl,tetrahydroquinolyl, isoquinolyl (such as, but not limited to, 1- and5-isoquinolyl), 1,2,3,4-tetrahydroisoquinolyl, cinnolinyl, quinoxalinyl(such as, but not limited to, 2- and 5-quinoxalinyl), quinazolinyl,phthalazinyl, 1,8-naphthyridinyl, 1,4-benzodioxanyl, coumarin,dihydrocoumarin, 1,5-naphthyridinyl, benzofuryl (such as, but notlimited to, 3-, 4-, 5-, 6- and 7-benzofuryl), 2,3-dihydrobenzofuryl,1,2-benzisoxazolyl, benzothienyl (such as, but not limited to, 3-, 4-,5-, 6-, and 7-benzothienyl), benzoxazolyl, benzothiazolyl (such as, butnot limited to, 2-benzothiazolyl and 5-benzothiazolyl), purinyl,benzimidazolyl, benztriazolyl, thioxanthinyl, carbazolyl, carbolinyl,acridinyl, pyrrolizidinyl, and quinolizidinyl.

The aforementioned listing of heterocyclyl and heteroaryl moieties isintended to be representative and not limiting.

As used herein, the term “substituted” means that an atom or group ofatoms has replaced hydrogen as the substituent attached to anothergroup.

For aryl, aryl-(C₁-C₃)alkyl and heterocyclyl groups, the term“substituted” as applied to the rings of these groups refers to anylevel of substitution, namely mono-, di-, tri-, tetra-, orpenta-substitution, where such substitution is permitted. Thesubstituents are independently selected, and substitution may be at anychemically accessible position. In one embodiment, the substituents varyin number between one and four. In another embodiment, the substituentsvary in number between one and three. In yet another embodiment, thesubstituents vary in number between one and two. In yet anotherembodiment, the substituents are independently selected from the groupconsisting of C₁₋₆ alkyl, —OH, C₁₋₆ alkoxy, halo, amino, acetamido andnitro. As used herein, where a substituent is an alkyl or alkoxy group,the carbon chain may be branched, straight or cyclic, in particular,straight.

“Instructional material,” as that term is used herein, includes apublication, a recording, a diagram, or any other medium of expressionthat can be used to communicate the usefulness of the composition and/orcompound of the invention in a kit. The instructional material of thekit may, for example, be affixed to a container that contains thecompound and/or composition of the invention or be shipped together witha container that contains the compound and/or composition.Alternatively, the instructional material may be shipped separately fromthe container with the intention that the recipient uses theinstructional material and the compound cooperatively. Delivery of theinstructional material may be, for example, by physical delivery of thepublication or other medium of expression communicating the usefulnessof the kit, or may alternatively be achieved by electronic transmission,for example by means of a computer, such as by electronic mail, ordownload from a website.

Throughout this disclosure, various aspects of the invention can bepresented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of theinvention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible sub-ranges as well asindividual numerical values within that range. For example, descriptionof a range such as from 1 to 6 should be considered to have specificallydisclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numberswithin that range, for example, 1, 2, 2.7, 3, 4, 5, 5.1, 5.3, 5.5, and6. This applies regardless of the breadth of the range.

Compounds and Compositions

The invention includes a compound of formula (I), or a salt or solvatethereof:

wherein in (I):R₁ is CR₅ or N, wherein:

-   -   if R₁ is CR₅, then R₃ is selected from the group consisting of        —N(R₅)₂, —NO, —N(R₅)N(R₅)₂, R₆, —N(R₅)—OR₅, —NH—C(═O)R₅, alkoxy,        —OSO₃H, —O(CR₅)_(n)R₆, —O(CR₅)_(n)alkoxy, —O(CR₅)_(n+1)OH,        —OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and        —OC(═O)C(R₅)═C(R₅)₂;

or R₃ is selected from the group consisting of ═O and ═S, and H* isomitted; and,

-   -   if R₁ is N, then R₃ is selected from the group consisting of        N(R₅)₂, —NO, —N(R₅)N(R₅)₂, R₆, —N(R₅)—OR₅, —NH—C(═O)R₅, Cl, Br,        I, alkoxy, mesyl, tosyl, —O(CR₅)_(n)R₆, —O(CR₅)_(n+1)OR₅,        —OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and        —OC(═O)C(R₅)═C(R₅)₂;        R₂ is selected from the group consisting of O, S, C(R₄)₂, and        N(R₄);        each occurrence of R₁ is independently selected from the group        consisting of H, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,        substituted alkynyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, substituted heteroarylalkyl, OR₅, and N(R₅)₂;        each occurrence of R₅ is independently selected from the group        consisting of H, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,        substituted alkynyl, aryl, substituted aryl, arylalkyl,        substituted arylalkyl, heteroaryl, substituted heteroaryl,        heteroarylalkyl, and substituted heteroarylalkyl;        R₆ is selected from the group consisting of F, Cl, Br, I, mesyl,        tosyl, —OSi(R₅)₃, —C(═O)OR₅, and —C(═O)R₅;        the dotted line is a single or double bond; and,        n is an integer ranging from 1 to 10.

In certain embodiments, the dotted line is a single bond. In otherembodiments, the dotted line is a double bond.

In certain embodiments, the compound of formula (I) is the compound offormula (Ia), or a salt or solvate thereof:

In certain embodiments, the compound of formula (I) is the compound offormula (Ib), or a salt or solvate thereof:

In certain embodiments, R₁ is N. In other embodiments, R₂ is N(R₄).

In certain embodiments, the compound of formula (I) is the compound offormula (Ic), or a salt or solvate thereof:

In certain embodiments, the compound of formula (I) is the compound offormula (Id), or a salt or solvate thereof:

In certain embodiments, R₃ is selected from the group consisting of—O(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and—OC(═O)C(R₅)═C(R₅)₂.

In certain embodiments, the compound of formula (I) is selected from thegroup consisting of:

In certain embodiments, R₁ is CR₅. In other embodiments, R₅ is selectedfrom the group consisting of H, alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, arylalkyl, substituted arylalkyl,heteroarylalkyl, and substituted heteroarylalkyl. In yet otherembodiments, R₃ is selected from the group consisting of R₆,—O(CR₅)_(n)R₆, OC(═O)(CR₅)_(n)R₇, and OC(═O)C(R₅)═C(R₅)₂; or R³ isselected from the group consisting of ═O and ═S, and H* is omitted.

In certain embodiments, n is 1, 2, 3, 4 or 5.

In certain embodiments, the salt is an acid addition salt and isselected from the group consisting of sulfate, hydrogen sulfate,hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric,phosphoric, formic, acetic, propionic, succinic, glycolic, gluconic,lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric,pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic,phenylacetic, mandelic, embonic (pamoic), methanesulfonic,ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic,2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic,cyclohexylaminosulfonic, stearic, alginic, -hydroxybutyric, salicylic,galactaric and galacturonic acid, and any combinations thereof.

In certain embodiments, the salt is a base addition salt and is selectedfrom the group consisting of calcium, magnesium, potassium, sodium,ammonium, zinc, a basic amine salt, and any combinations thereof,wherein the basic amine is selected from the group consisting oftriethylamine, diisopropylethylamine, trimethylamine, N,N′-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine, procaine and any combinations thereof.

The compounds of the invention may possess one or more stereocenters,and each stereocenter may exist independently in either the (R) or (S)configuration. In one embodiment, compounds described herein are presentin optically active or racemic forms. The compounds described hereinencompass racemic, optically-active, regioisomeric and stereoisomericforms, or combinations thereof that possess the therapeutically usefulproperties described herein. Preparation of optically active forms isachieved in any suitable manner, including by way of non-limitingexample, by resolution of the racemic form with recrystallizationtechniques, synthesis from optically-active starting materials, chiralsynthesis, or chromatographic separation using a chiral stationaryphase. In one embodiment, a mixture of one or more isomer is utilized asthe therapeutic compound described herein. In another embodiment,compounds described herein contain one or more chiral centers. Thesecompounds are prepared by any means, including stereoselectivesynthesis, enantioselective synthesis and/or separation of a mixture ofenantiomers and/or diastereomers. Resolution of compounds and isomersthereof is achieved by any means including, by way of non-limitingexample, chemical processes, enzymatic processes, fractionalcrystallization, distillation, and chromatography.

The methods and formulations described herein include the use ofN-oxides (if appropriate), crystalline forms (also known as polymorphs),solvates, amorphous phases, and/or pharmaceutically acceptable salts ofcompounds having the structure of any compound of the invention, as wellas metabolites and active metabolites of these compounds having the sametype of activity. Solvates include water, ether (e.g., tetrahydrofuran,methyl tert-butyl ether) or alcohol (e.g., ethanol) solvates, acetatesand the like. In one embodiment, the compounds described herein exist insolvated forms with pharmaceutically acceptable solvents such as water,and ethanol. In another embodiment, the compounds described herein existin unsolvated form.

In one embodiment, the compounds of the invention exist as tautomers.All tautomers are included within the scope of the compounds recitedherein.

In one embodiment, compounds described herein are prepared as prodrugs(see for example Hacker, et al., Pharmacology: Principles and Practice.Academic Press, Jun 19, 2009. pp. 216-217). A “prodrug” is an agentconverted into the parent drug in vivo. In one embodiment, upon in vivoadministration, a prodrug is chemically converted to the biologically,pharmaceutically or therapeutically active form of the compound. Inanother embodiment, a prodrug is enzymatically metabolized by one ormore steps or processes to the biologically, pharmaceutically ortherapeutically active form of the compound.

In certain embodiments, sites on, for example, the aromatic ring portionof compounds of the invention are susceptible to various metabolicreactions. Incorporation of appropriate substituents on the aromaticring structures may reduce, minimize or eliminate this metabolicpathway. Accordingly, in one embodiment, a prodrug is created by methodswell known in the art, by which the appropriate substituent to decreaseor eliminate the susceptibility of the aromatic ring to metabolicreactions is added, by way of example only, a deuterium, a halogen, oran alkyl group.

Compounds described herein also include isotopically-labeled compoundswherein one or more atoms is replaced by an atom having the same atomicnumber, but an atomic mass or mass number different from the atomic massor mass number usually found in nature. Examples of isotopes suitablefor inclusion in the compounds described herein include and are notlimited to ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ³⁶Cl, ¹⁸F, ¹²³I, ¹²⁵I, ¹³N, ¹⁵N, ¹⁵O,¹⁷O, ¹⁸O, ³²P, and ³⁵S. In one embodiment, isotopically-labeledcompounds are useful in drug and/or substrate tissue distributionstudies. In another embodiment, substitution with heavier isotopes suchas deuterium affords greater metabolic stability (for example, increasedin vivo half-life or reduced dosage requirements). In yet anotherembodiment, substitution with positron emitting isotopes, such as ¹¹C,¹⁸F, ¹⁵O and ¹³N, is useful in Positron Emission Topography (PET)studies for examining substrate receptor occupancy. Isotopically-labeledcompounds are prepared by any suitable method or by processes using anappropriate isotopically-labeled reagent in place of the non-labeledreagent otherwise employed.

In one embodiment, the compounds described herein are labeled by othermeans, including, but not limited to, the use of chromophores orfluorescent moieties, bioluminescent labels, or chemiluminescent labels.

The invention further includes a pharmaceutical composition comprisingthe compound of the invention and a pharmaceutically acceptable carrier.

In certain embodiments, the pharmaceutical composition further comprisesat least one additional chemotherapeutic agent selected from the groupconsisting of alkylating agents; nitrosoureas; antimetabolites;antitumor antibiotics; plant alkyloids; taxanes; hormonal agents;anti-angiogenesis agents, and miscellaneous agents.

In certain embodiments, the pharmaceutical composition further comprisesat least one additional anti-angiogenesis agent. In other embodiments,the anti-angiogenesis agent is at least one selected from the groupconsisting of 2-methoxyestradiol AG3340, angiostatin, antithrombin-III,anti-VEGF antibody, VEGF antagonist, batimastat, bevacizumab,BMS-275291, CA1, canstatin, combretastatin, comhretastatin-A4 phosphate,CC-5013, captopril, celecoxib, dalteparin, EMD121974, endostatin,erlotinib, gefitinib, genistein, halofuginone, ID1, ID3, IM862, omatinibmesylate, inducible protein-10, interferon-alpha, interleukin-12,lavendustin-a, LY317615, AE-941, merimastat, mapsin, medroxpregesteronacetate, Meth-1, Meth-2, Neovastat, osteopontin cleaved product, PEX,pigment epithelium growth factor, platelet growth factor 4, prolactinfragment, proliferin-related protein (PRP), PTK787 /ZK222584,recombinant human platelet factor-4, restin, squalamine, SU5416, SU6668,suramin, taxol, tecogalan, thalidomide, thrombospondin, TNP-470,troponin I, vasostatin, VEGF1, VEGF-TRAP and ZD6474.

In certain embodiments, the compound of the invention and the additionalagent are co-formulated in the composition.

Synthesis

The compounds described herein, and other related compounds havingdifferent substituents are synthesized using techniques and materialsdescribed herein and as described, for example, in Fieser & Fieser'sReagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons,1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 andSupplementals (Elsevier Science Publishers, 1989); Organic Reactions,Volumes 1-40 (John Wiley and Sons, 1991), Larock's Comprehensive OrganicTransformations (VCH Publishers Inc., 1989), March, Advanced OrganicChemistry 4^(th) Ed., (Wiley 1992); Carey & Sundberg, Advanced OrganicChemistry 4th Ed., Vols. A and B (Plenum 2000, 2001), and Green & Wuts,Protective Groups in Organic Synthesis 3rd Ed., (Wiley 1999) (all ofwhich are incorporated by reference for such disclosure). Generalmethods for the preparation of compound as described herein are modifiedby the use of appropriate reagents and conditions, for the introductionof the various moieties found in the formula as provided herein.

Compounds described herein are synthesized using any suitable proceduresstarting from compounds that are available from commercial sources, orare prepared using procedures described herein.

In one embodiment, reactive functional groups, such as hydroxyl, amino,imino, thio or carboxy groups, are protected in order to avoid theirunwanted participation in reactions. Protecting groups are used to blocksome or all of the reactive moieties and prevent such groups fromparticipating in chemical reactions until the protective group isremoved. In another embodiment, each protective group is removable by adifferent means. Protective groups that are cleaved under totallydisparate reaction conditions fulfill the requirement of differentialremoval.

In one embodiment, protective groups are removed by acid, base, reducingconditions (such as, for example, hydrogenolysis), and/or oxidativeconditions. Groups such as trityl, dimethoxytrityl, acetal andt-butyldimethylsilyl are acid labile and are used to protect carboxy andhydroxy reactive moieties in the presence of amino groups protected withCbz groups, which are removable by hydrogenolysis, and Fmoc groups,which are base labile. Carboxylic acid and hydroxy reactive moieties areblocked with base labile groups such as, but not limited to, methyl,ethyl, and acetyl, in the presence of amines that are blocked with acidlabile groups, such as t-butyl carbamate, or with carbamates that areboth acid and base stable but hydrolytically removable.

In one embodiment, carboxylic acid and hydroxy reactive moieties areblocked with hydrolytically removable protective groups such as thebenzyl group, while amine groups capable of hydrogen bonding with acidsare blocked with base labile groups such as Fmoc. Carboxylic acidreactive moieties are protected by conversion to simple ester compoundsas exemplified herein, which include conversion to alkyl esters, or areblocked with oxidatively-removable protective groups such as2,4-dimethoxy benzyl, while co-existing amino groups are blocked withfluoride labile silyl carbamates.

Ally! blocking groups are useful in the presence of acid- andbase-protecting groups since the former are stable and are subsequentlyremoved by metal or pi-acid catalysts. For example, an allyl-blockedcarboxylic acid is deprotected with a palladium-catalyzed reaction inthe presence of acid labile t-butyl carbamate or base-labile acetateamine protecting groups. Yet another form of protecting group is a resinto which a compound or intermediate is attached. As long as the residueis attached to the resin, that functional group is blocked and does notreact. Once released from the resin, the functional group is availableto react.

Typically blocking/protecting groups may be selected from:

Other protecting groups, plus a detailed description of techniquesapplicable to the creation of protecting groups and their removal aredescribed in Greene & Wuts, Protective Groups in Organic Synthesis, 3rdEd., John Wiley & Sons, New York, N.Y., 1999, and Kocienski, ProtectiveGroups, Thieme Verlag, New York, N.Y., 1994, which are incorporatedherein by reference for such disclosure.

The compounds of the invention may be prepared according to the generalmethodology illustrated in the synthetic schemes described below. Thereagents and conditions described herein may be modified to allow thepreparation of the compounds of the invention, and such modificationsare known to those skilled in the art. The scheme included herein areintended to illustrate but not limit the chemistry and methodologiesthat one skilled in the art may use to make compounds of the invention.

In certain embodiments, the compounds of the invention, or intermediatesthat are useful in preparing the compounds of the invention, may begenerated using a Diels-Alder reaction between 7-hydrocholesterol, or aderivative thereof, such as compound (II), with a five-membereddienophile, such as compound (III):

The product of the Diels-Alder reaction may be the compound of theinvention, or may be further derivatized to yield a compound of theinvention, as will be appreciated by those skilled in the art.

Salts

The compounds described herein may form salts with acids, and such saltsare included in the present invention. In one embodiment, the salts arepharmaceutically acceptable salts. The term “salts” embraces additionsalts of free acids that are useful within the methods of the invention.The term “pharmaceutically acceptable salt” refers to salts that possesstoxicity profiles within a range that affords utility in pharmaceuticalapplications. Pharmaceutically unacceptable salts may nonethelesspossess properties such as high crystallinity, which have utility in thepractice of the present invention, such as for example utility inprocess of synthesis, purification or formulation of compounds usefulwithin the methods of the invention.

Suitable pharmaceutically acceptable acid addition salts may be preparedfrom an inorganic acid or from an organic acid. Examples of inorganicacids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic,sulfuric (including sulfate and hydrogen sulfate), and phosphoric acids(including hydrogen phosphate and dihydrogen phosphate). Appropriateorganic acids may be selected from aliphatic, cycloaliphatic, aromatic,araliphatic, heterocyclic, carboxylic and sulfonic classes of organicacids, examples of which include formic, acetic, propionic, succinic,glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic,glucuronic, maleic, malonic, saccharin, fumaric, pyruvic, aspartic,glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic,mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic,benzenesulfonic, pantothenic, trifluoromethanesulfonic,2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic,cyclohexylaminosulfonic, stearic, alginic, -hydroxybutyric, salicylic,galactaric and galacturonic acid.

Suitable pharmaceutically acceptable base addition salts of compounds ofthe invention include, for example, metallic salts including alkalimetal, alkaline earth metal and transition metal salts such as, forexample, calcium, magnesium, potassium, sodium and zinc salts.Pharmaceutically acceptable base addition salts also include organicsalts made from basic amines such as, for example, N,N′-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine (N-methylglucamine) and procaine. All ofthese salts may be prepared from the corresponding compound by reacting,for example, the appropriate acid or base with the compound.

Methods

The invention includes a method of preventing or treating cancer in asubject in need thereof. The invention further includes a method ofpreventing, reversing or inhibiting angiogenesis in a subject in needthereof. The invention further includes a method of reducing obesity andobesity-induced disorders, such as diabetes and cardiovascular diseases,in a subject in need thereof. The invention further includes a method oftreating cholesterol disorders in a subject in need thereof. Theinvention further includes a method of treatring a patient sufferingfrom Smith-Lemli-Opitz syndrome (SLOS). The invention further includes amethod of diagnosing a disease or disorder in a subject in need thereof.The invention further includes a method of inhibiting activity of thevitamin D nuclear receptor. The invention further includes a method ofinhibiting the activity of the vitamin D nuclear receptor (also known ascalcitriol receptor) in a subject in need thereof.

In certain embodiments, the method comprises administering to thesubject a therapeutically effective amount of at least one compound offormula (I) or a salt or solvate thereof:

wherein in (I):

R₁ is CR₅ or N;

R₃ is selected from the group consisting of —N(R₅)₂, —NO, —N(R₅)N(R₅)₂,R₆, —N(R₅)—OR₅, —NH—C(═O)R₅, F, Cl, Br, I, hydroxy, alkoxy, mesyl,tosyl, —OSO₃H, —O(CR₅)_(n)R₆, —O(CR₅)_(n)alkoxy, —O(CR₅)_(n+1)OH,—OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and —OC(═O)C(R₅)═C(R₅)₂;

or R₃ is selected from the group consisting of ═O and ═S, and H* isomitted;

R₂ is selected from the group consisting of O, S, C(R₄)₂, and N(R₄);

each occurrence of R₄ is independently selected from the groupconsisting of H, alkyl, substituted alkyl, cycloalkyl, substitutedcycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl,substituted heteroaryl, heteroarylalkyl, substituted heteroarylalkyl,OR₅, and N(R₅)₂; each occurrence of R₅ is independently selected fromthe group consisting of H, alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, andsubstituted heteroarylalkyl;

R₆ is selected from the group consisting of F, Cl, Br, I, mesyl, tosyl,—OSi(R₅)₃, —C(═O)OR₅, and —C(═O)R₅;

the dotted line is a single or double bond; and,

n is an integer ranging from 1 to 10.

In certain embodiments, the method comprises contacting the vitamin Dnuclear receptor with an effective amount of at least one compound offormula (I) or a salt or solvate thereof.

In certain embodiments, if R₁ is CR₅, then R₃ is selected from the groupconsisting of —N(R₅)₂, —NO, —N(R₅)N(R₅)₂, R₆, —N(R₅)—OR₅, —NH—C(═O)R₅,alkoxy, —OSO₃H, —O(CR₅)_(n)R₆, —O(CR₅)_(n)alkoxy, —O(CR₅)_(n+1)OH,—OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and —OC(═O)C(R₅)═C(R₅)₂; or R₃is selected from the group consisting of ═O and ═S, and H* is omitted.

In certain embodiments, if R₁ is N, then R₃ is selected from the groupconsisting of N(R₅)₂, —NO, —N(R₅)N(R₅)₂, R₆, —N(R₅)—OR₅, —NII—C(═O)R₅,Cl, Br, I, alkoxy, mesyl, tosyl, —O(CR5)_(n)R₆, —O(CR₅)_(n+1)OR₅,—OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and —OC(═O)C(R₅)═C(R₅)₂.

In certain embodiments, the compound of formula (I) is the compound offormula (Ia) or a salt or solvate thereof:

In certain embodiments, the compound of formula (I) is the compound offormula (Ib), or a salt or solvate thereof:

In certain embodiments, R₁ is N. In other embodiments, R₂ is N(R₄).

In certain embodiments, the compound of formula (I) is the compound offormula (Ic), or a salt or solvate thereof:

In certain embodiments, the compound of formula (1) is the compound offoimula (Id), or a salt or solvate thereof:

In certain embodiments, R₃ is selected from the group consisting of—O(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and—OC(═O)C(R₅)═C(R₅)₂.

In certain embodiments, the compound of formula (I) is selected from thegroup consisting of:

In certain embodiments, the cancer comprises at least one selected fromthe group consisting of breast cancer, prostate cancer, ovarian cancer,cervical cancer, skin cancer, pancreatic cancer, colorectal cancer,renal cancer, liver cancer, brain cancer, lymphoma, leukemia, lungcancer, endometrial cancer, neuroblastoma, and any combinations thereof.In other embodiments, the cancer comprises at least one selected fromthe group consisting of prostate cancer, breast cancer, ovarian cancer,endometrial cancer, medulloblastoma, neuroblastoma, melanoma, and anycombinations thereof.

In certain embodiments, the subject is further administered at least oneadditional chemotherapeutic agent. In other embodiments, thechemotherapeutic agent is selected from the group consisting ofalkylating agents; nitrosoureas; antimetabolites; antitumor antibiotics;plant alkyloids; taxanes; hormonal agents; anti-angiogenesis agents, andmiscellaneous agents.

In certain embodiments, the subject is further administered at least oneadditional anti-angiogenesis agent. In other embodiments, theanti-angiogenesis agent is at least one selected from the groupconsisting of 2-methoxyestradiol AG3340, angiostatin, antithrombin-III,anti-VEGF antibody, VEGF antagonist, batimastat, bevacizumab,BMS-275291, CA1, canstatin, combretastatin, combretastatin-A4 phosphate,CC-5013, captopril, celecoxib, dalteparin, EMD121974, endostatin,erlotinib, gefitinib, genistein, halofuginone, IDE ID3, IM862, omatinibmesylate, inducible protein-10, interferon-alpha, interleukin-12,lavcndustin-a, LY317615, AE-941, merimastat, mapsin, medroxpregesteronacetate, Meth-1, Meth-2, Neovastat, osteopontin cleaved product, PEX,pigment epithelium growth factor, platelet growth factor 4, prolactinfragment, proliferin-related protein (PRP), PTK787 /ZK222584,recombinant human platelet factor-4, restin, squalamine, SU5416, SU6668,suramin, taxol, tecogalan, thalidomide, thrombospondin, TNP-470,troponin I, vasostatin, VEGF1, VEGF-TRAP and ZD6474.

In certain embodiments, the compound of the invention and the additionalagent are separately administered to the subject. In other embodiments,the compound of the invention and the additional agent areco-administered to the subject. In yet other embodiments, the compoundof the invention and the additional agent are co-formulated.

In certain embodiments, the subject is a mammal. In other embodiments,the mammal is a human.

In certain embodiments, the composition is administered to the subjectby at least one route selected from the group consisting of intravenous,oral, inhalational, rectal, vaginal, transdermal, intranasal, buccal,sublingual, parenteral, intrathecal, intragastrical, ophthalmic,pulmonary and topical routes.

In certain embodiments, the method further comprises procuring thecompound of the invention for the subject.

Kits

The invention includes a kit comprising a compound of the invention, anapplicator, and an instructional material for use thereof. Theinstructional material included in the kit comprises instructions forpreventing or treating a disorder or disease contemplated within theinvention in a subject. The instructional material recites the amountof, and frequency with which, the compound of the invention should beadministered to the subject. In certain embodiments, the kit furthercomprises at least one additional chemotherapeutic agent. In otherembodiments, the kit further comprises at least one additionalanti-angiogenesis agent.

Combination Therapies

In certain embodiments, the compounds of the invention are useful in themethods of the invention in combination with at least one additionalcompound useful for treating or preventing cancer and/or uncontrolledangiogenesis. This additional compound may comprise compounds identifiedherein or compounds, e.g., commercially available compounds, known totreat, prevent or reduce the symptoms of cancer and/or uncontrolledangiogenesis.

In one aspect, the present invention contemplates that the compounds ofthe invention may be used in combination with a therapeutic agent suchas an antitumor agent, including but not limited to a chemotherapeuticagent, an anti-cell proliferation agent or any combination thereof. Forexample, any conventional chemotherapeutic agents of the followingnon-limiting exemplary classes are included in the invention: alkylatingagents; nitrosoureas; antimetabolites; antitumor antibiotics; plantalkyloids; taxanes; hormonal agents; anti-angiogenesis agents, andmiscellaneous agents.

Alkylating agents are so named because of their ability to add alkylgroups to many electronegative groups under conditions present in cells,thereby interfering with DNA replication to prevent cancer cells fromreproducing. Most alkylating agents are cell cycle non-specific. Inspecific aspects, they stop tumor growth by cross-linking guanine basesin DNA double-helix strands. Non-limiting examples include busulfan,carboplatin, chlorambucil, cisplatin, cyclophosphamide, dacarbazine,ifosfamide, mechlorethamine hydrochloride, melphalan, procarbazine,thiotepa, and uracil mustard.

Anti-metabolites prevent incorporation of bases into DNA during thesynthesis (S) phase of the cell cycle, prohibiting normal developmentand division. Non-limiting examples of antimetabolites include drugssuch as 5-fluorouracil, 6-mercaptopurine, capecitabine, cytosinearabinoside, floxuridine, fludarabine, gemcitabine, methotrexate, andthioguanine.

Antitumor antibiotics generally prevent cell division by interferingwith enzymes needed for cell division or by altering the membranes thatsurround cells. Included in this class are the anthracyclines, such asdoxorubicin, which act to prevent cell division by disrupting thestructure of the DNA and terminate its function. These agents are cellcycle non-specific. Non-limiting examples of antitumor antibioticsinclude dactinomycin, daunorubicin, doxorubicin, idarubicin,mitomycin-C, and mitoxantrone.

Plant alkaloids inhibit or stop mitosis or inhibit enzymes that preventcells from making proteins needed for cell growth. Frequently used plantalkaloids include vinblastine, vincristine, vindesine, and vinorelbine.However, the invention should not be construed as being limited solelyto these plant alkaloids.

The taxanes affect cell structures called microtubules that areimportant in cellular functions. In normal cell growth, microtubules areformed when a cell starts dividing, but once the cell stops dividing,the microtubules are disassembled or destroyed. Taxanes prohibit themicrotubules from breaking down such that the cancer cells become soclogged with microtubules that they cannot grow and divide. Non-limitingexemplary taxanes include paclitaxel and docetaxel.

Hormonal agents and hormone-like drugs are utilized for certain types ofcancer, including, for example, leukemia, lymphoma, and multiplemyeloma. They are often employed with other types of chemotherapy drugsto enhance their effectiveness. Sex hormones are used to alter theaction or production of female or male hormones and are used to slow thegrowth of breast, prostate, and endometrial cancers. Inhibiting theproduction (aromatase inhibitors) or action (tamoxifen) of thesehormones can often be used as an adjunct to therapy. Some other tumorsare also hormone dependent. Tamoxifen is a non-limiting example of ahormonal agent that interferes with the activity of estrogen, whichpromotes the growth of breast cancer cells.

Examples of anti-angiogenesis agents include but are not limited to2-methoxyestradiol (2-ME), AG3340, angiostatin, antithrombin-III,anti-VEGF antibody, batimastat, bevacizumab (Avastin), BMS-275291, CAI,canstatin, combretastatin, combretastatin-A4 phosphate, CC-5013,captopril, celecoxib, daltcparin, EMD121974, endostatin, erlotinib,gefitinib, genistein, halofuginone, ID1, ID3, IM862, omatinib mesylate,inducible protein-10, interferon-alpha, interleukin-12, lavendustin-a,LY317615, AE-941, merimastat, mapsin, medroxpregesteron acetate, Meth-1,Meth-2, Neovastat, osteopontin cleaved product, PEX, pigment epitheliumgrowth factor (PEGF), platelet growth factor 4, prolactin fragment,proliferin-related protein(PRP), PTK787 /ZK222584, recombinant humanplatelet factor-4 (rPF4), restin, squalamine, SU5416, SU6668, suramin,taxol, tecogalan, thalidomide, thrombospondin, TNP-470, troponin I,vasostatin, VEGF1, VEGF-TRAP and ZD6474. In some embodiments theanti-angiogenesis agent is a VEGF antagonist, such as a VEGF bindingmolecule (such as VEGF antibodies, or antigen binding fragment(s)thereof) or a VEGF antagonist such as NeXstar.

Miscellaneous agents include chemotherapeutics such as bleomycin,hydroxyurea, L-asparaginase, and procarbazine that are also useful inthe invention.

An anti-cell proliferation agent can further be defined as anapoptosis-inducing agent or a cytotoxic agent. The apoptosis-inducingagent may be a granzyme, a Bcl-2 family member, cytochrome C, a caspase,or a combination thereof. Exemplary granzymes include granzyme A,granzyme B, granzyme C, granzyme D, granzyme E, granzyme F, granzyme G,granzyme H, granzyme I, granzyme J, granzyme K, granzyme L, granzyme M,granzyme N, or a combination thereof. In other specific aspects, theBcl-2 family member is, for example, Bax, Bak, Bcl-Xs, Bad, Bid. Bik,Hrk, Bok, or a combination thereof.

In additional aspects, the caspase is caspase-1, caspase-2, caspase-3,caspase-4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9,caspase-10, caspase-11, caspase-12, caspase-13, caspase-14, or acombination thereof. In specific aspects, the cytotoxic agent is TNF-α,gelonin, Prodigiosin, a ribosome-inhibiting protein (RIP). Pseudomonasexotoxin, Clostridium difficile Toxin B, Helicobacter pylori VacA,Yersinia enterocolitica YopT, Violacein, diethylenetriaminepentaaceticacid, irofulven, Diptheria Toxin, mitogillin, ricin, botulinum toxin,cholera toxin, saporin 6, or a combination thereof.

A synergistic effect may be calculated, for example, using suitablemethods such as, for example, the Sigmoid-E_(max) equation (Holford &Scheiner, 19981, Clin. Pharmacokinet. 6: 429-453), the equation of Loeweadditivity (Loewe & Muischnek, 1926, Arch. Exp. Pathol Pharmacol. 114:313-326) and the median-effect equation (Chou & Talalay, 1984, Adv.Enzyme Regul. 22:27-55). Each equation referred to above may be appliedto experimental data to generate a corresponding graph to aid inassessing the effects of the drug combination. The corresponding graphsassociated with the equations referred to above are theconcentration-effect curve, isobologram curve and combination indexcurve, respectively.

Administration/Dosage/Formulations

The regimen of administration may affect what constitutes an effectiveamount. The therapeutic formulations may be administered to the subjecteither prior to or after the onset of a disease or disorder contemplatedin the invention. Further, several divided dosages, as well as staggereddosages may be administered daily or sequentially, or the dose may becontinuously infused, or may be a bolus injection. Further, the dosagesof the therapeutic formulations may be proportionally increased ordecreased as indicated by the exigencies of the therapeutic orprophylactic situation.

Administration of the compositions of the present invention to apatient, in particular a mammal, more particularly a human, may becarried out using known procedures, at dosages and for periods of timeeffective to treat a disease or disorder contemplated in the invention.An effective amount of the therapeutic compound necessary to achieve atherapeutic effect may vary according to factors such as the state ofthe disease or disorder in the patient; the age, sex, and weight of thepatient; and the ability of the therapeutic compound to treat a diseaseor disorder contemplated in the invention. Dosage regimens may beadjusted to provide the optimum therapeutic response. For example,several divided doses may be administered daily or the dose may beproportionally reduced as indicated by the exigencies of the therapeuticsituation. A non-limiting example of an effective dose range for atherapeutic compound of the invention is from about 1 and 5,000 mg/kg ofbody weight/per day. One of ordinary skill in the art would be able tostudy the relevant factors and make the determination regarding theeffective amount of the therapeutic compound without undueexperimentation.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

In particular, the selected dosage level depends upon a variety offactors including the activity of the particular compound employed, thetime of administration, the rate of excretion of the compound, theduration of the treatment, other drugs, compounds or materials used incombination with the compound, the age, sex, weight, condition, generalhealth and prior medical history of the patient being treated, and likefactors well, known in the medical arts.

A medical doctor, e.g., physician or veterinarian, having ordinary skillin the art may readily determine and prescribe the effective amount ofthe pharmaceutical composition required. For example, the physician orveterinarian could start doses of the compounds of the inventionemployed in the pharmaceutical composition at levels lower than thatrequired in order to achieve the desired therapeutic effect andgradually increase the dosage until the desired effect is achieved.

In particular embodiments, it is especially advantageous to formulatethe compound in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the patients tobe treated; each unit containing a predetermined quantity of therapeuticcompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical vehicle. The dosage unitforms of the invention are dictated by and directly dependent on (a) theunique characteristics of the therapeutic compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding/formulating such a therapeutic compound for thetreatment of a disease or disorder contemplated in the invention.

In one embodiment, the compositions of the invention are formulatedusing one or more pharmaceutically acceptable excipients or carriers. Inone embodiment, the pharmaceutical compositions of the inventioncomprise a therapeutically effective amount of a compound of theinvention and a pharmaceutically acceptable carrier.

The carrier may be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), suitable mixturesthereof, and vegetable oils. The proper fluidity may be maintained, forexample, by the use of a coating such as lecithin, by the maintenance ofthe required particle size in the case of dispersion and by the use ofsurfactants. Prevention of the action of microorganisms may be achievedby various antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it is advantageous to include isotonic agents, for example,sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol,in the composition. Prolonged absorption of the injectable compositionsmay be brought about by including in the composition an agent whichdelays absorption, for example, aluminum monostearate or gelatin.

In one embodiment, the compounds/compositions of the invention areadministered to the patient in dosages that range from one to five timesper day or more. In another embodiment, the compositions of theinvention are administered to the patient in range of dosages thatinclude, but are not limited to, once every day, every two, days, everythree days to once a week, and once every two weeks. It is readilyapparent to one skilled in the art that the frequency of administrationof the various combination compositions of the invention varies fromindividual to individual depending on many factors including, but notlimited to, age, disease or disorder to be treated, gender, overallhealth, and other factors. Thus, the invention should not be construedto be limited to any particular dosage regime and the precise dosage andcomposition to be administered to any patient is determined by theattending physical taking all other factors about the patient intoaccount.

Compounds of the invention for administration may be in the range offrom about 1 μg to about 10,000 mg, about 20 μg to about 9,500 mg, about40 μg to about 9,000 mg. about 75 μg to about 8,500 mg, about 150 μg toabout 7,500 mg, about 200 μg to about 7,000 mg, about 3050 μg to about6,000 mg, about 500 μg to about 5,000 mg, about 750 μg to about 4,000mg, about 1 mg to about 3,000 mg, about 10 mg to about 2,500 mg, about20 mg to about 2,000 mg, about 25 mg to about 1,500 mg, about 30 mg toabout 1,000 mg, about 40 mg to about 900 mg, about 50 mg to about 800mg, about 60 mg to about 750 mg, about 70 mg to about 600 mg, about 80mg to about 500 mg, and any and all whole or partial incrementstherebetween.

In some embodiments, the dose of a compound of the invention is fromabout 1 mg and about 2,500 mg. In some embodiments, a dose of a compoundof the invention used in compositions described herein is less thanabout 10,000 mg, or less than about 8,000 mg, or less than about 6,000mg, or less than about 5,000 mg, or less than about 3,000 mg, or lessthan about 2,000 mg, or less than about 1,000 mg, or less than about 500mg, or less than about 200 mg, or less than about 50 mg. Similarly, insome embodiments, a dose of a second compound as described herein isless than about 1,000 mg, or less than about 800 mg, or less than about600 mg, or less than about 500 mg. or less than about 400 mg, or lessthan about 300 mg, or less than about 200 mg, or less than about 100 mg,or less than about 50 mg, or less than about 40 mg, or less than about30 mg, or less than about 25 mg, or less than about 20 mg, or less thanabout 15 mg, or less than about 10 mg, or less than about 5 mg, or lessthan about 2 mg, or less than about 1 mg, or less than about 0.5 mg, andany and all whole or partial increments thereof.

In one embodiment, the present invention is directed to a packagedpharmaceutical composition comprising a container holding atherapeutically effective amount of a compound of the invention, aloneor in combination with a second pharmaceutical agent; and instructionsfor using the compound to treat, prevent, or reduce one or more symptomsof a disease or disorder contemplated in the invention.

Formulations may be employed in admixtures with conventional excipients,i.e., pharmaceutically acceptable organic or inorganic carriersubstances suitable for oral, parenteral, nasal, intravenous,subcutaneous, enteral. or any other suitable mode of administration,known to the art. The pharmaceutical preparations may be sterilized andif desired mixed with auxiliary agents, e.g., lubricants, preservatives,stabilizers, wetting agents, emulsifiers, salts for influencing osmoticpressure buffers, coloring, flavoring and/or aromatic substances and thelike. They may also be combined where desired with other active agents,e.g., other analgesic agents.

Routes of administration of any of the compositions of the inventioninclude oral, nasal, rectal, intravaginal, parenteral, buccal,sublingual or topical. The compounds for use in the invention may beformulated for administration by any suitable route, such as for oral orparenteral, for example, transdermal, transmucosal (e.g., sublingual,lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- andperivaginally), (intra)nasal and (trans)rectal), intravesical,intrapulmonary, intraduodenal, intragastrical, intrathecal,subcutaneous, intramuscular, intradermal, intra-arterial, intravenous,intrabronchial, inhalation, and topical administration.

Suitable compositions and dosage forms include, for example, tablets,capsules, caplets, pills, gel caps, troches, dispersions, suspensions,solutions, syrups, granules, beads, transdermal patches, gels, powders,pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs,suppositories, liquid sprays for nasal or oral administration, drypowder or aerosolized formulations for inhalation, compositions andformulations for intravesical administration and the like. It should beunderstood that the formulations and compositions that would be usefulin the present invention are not limited to the particular formulationsand compositions that are described herein.

Oral Administration

For oral application, particularly suitable are tablets, dragees,liquids, drops, suppositories, or capsules, caplets and gelcaps. Thecompositions intended for oral use may be prepared according to anymethod known in the art and such compositions may contain one or moreagents selected from the group consisting of inert, non-toxicpharmaceutically excipients that are suitable for the manufacture oftablets. Such excipients include, for example an inert diluent such aslactose; granulating and disintegrating agents such as cornstarch;binding agents such as starch; and lubricating agents such as magnesiumstearate. The tablets may be uncoated or they may be coated by knowntechniques for elegance or to delay the release of the activeingredients. Formulations for oral use may also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertdiluent.

For oral administration, the compounds of the invention may be in theform of tablets or capsules prepared by conventional means withpharmaceutically acceptable excipients such as binding agents (e.g.,polyvinylpyrrolidone, hydroxypropylcellulose orhydroxypropylmethylcellulose); fillers (e.g., cornstarch, lactose,microcrystalline cellulose or calcium phosphate); lubricants (e.g.,magnesium stearate, talc, or silica); disintegrates (e.g., sodium starchglycollate); or wetting agents (e.g., sodium lauryl sulphate). Ifdesired, the tablets may be coated using suitable methods and coatingmaterials such as OPADRY™ film coating systems available from Colorcon,West Point, Pa. (e.g., OPADRY™ OY Type, OYC Type, Organic Enteric OY-PType, Aqueous Enteric OY-A Type, OY-PM Type and OPADRY™ White,32K18400). Liquid preparation for oral administration may be in the formof solutions, syrups or suspensions. The liquid preparations may beprepared by conventional means with pharmaceutically acceptableadditives such as suspending agents (e.g., sorbitol syrup, methylcellulose or hydrogenated edible fats); emulsifying agent (e.g.,lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily estersor ethyl alcohol); and preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid).

Granulating techniques are well known in the pharmaceutical art formodifying starting powders or other particulate materials of an activeingredient. The powders are typically mixed with a binder material intolarger permanent free-flowing agglomerates or granules referred to as a“granulation.” For example, solvent-using “wet” granulation processesare generally characterized in that the powders are combined with abinder material and moistened with water or an organic solvent underconditions resulting in the formation of a wet granulated mass fromwhich the solvent must then be evaporated.

Melt granulation generally consists in the use of materials that aresolid or semi-solid at room temperature (i.e. having a relatively lowsoftening or melting point range) to promote granulation of powdered orother materials, essentially in the absence of added water or otherliquid solvents. The low melting solids, when heated to a temperature inthe melting point range, liquefy to act as a binder or granulatingmedium. The liquefied solid spreads itself over the surface of powderedmaterials with which it is contacted, and on cooling, forms a solidgranulated mass in which the initial materials are bound together. Theresulting melt granulation may then be provided to a tablet press or beencapsulated for preparing the oral dosage form. Melt granulationimproves the dissolution rate and bioavailability of an active (i.e.drug) by forming a solid dispersion or solid solution.

U.S. Pat. No. 5,169,645 discloses directly compressible wax-containinggranules having improved flow properties. The granules are obtained whenwaxes are admixed in the melt with certain flow improving additives,followed by cooling and granulation of the admixture. In certainembodiments, only the wax itself melts in the melt combination of thewax(es) and additives(s), and in other cases both the wax(es) and theadditives(s) melt.

The present invention also includes a multi-layer tablet comprising alayer providing for the delayed release of one or more compounds of theinvention, and a further layer providing for the immediate release of amedication for treatment of a disease or disorder contemplated in theinvention. Using a wax/pH-sensitive polymer mix, a gastric insolublecomposition may be obtained in which the active ingredient is entrapped,ensuring its delayed release.

Parenteral Administration

As used herein, “parenteral administration” of a pharmaceuticalcomposition includes any route of administration characterized byphysical breaching of a tissue of a subject and administration of thepharmaceutical composition through the breach in the tissue. Parenteraladministration thus includes, but is not limited to, administration of apharmaceutical composition by injection of the composition, byapplication of the composition through a surgical incision, byapplication of the composition through a tissue-penetrating non-surgicalwound, and the like. In particular, parenteral administration iscontemplated to include, but is not limited to, subcutaneous,intravenous, intraperitoneal, intramuscular, intrasternal injection, andkidney dialytic infusion techniques.

Formulations of a pharmaceutical composition suitable for parenteraladministration comprise the active ingredient combined with apharmaceutically acceptable carrier, such as sterile water or sterileisotonic saline. Such formulations may be prepared, packaged, or sold ina form suitable for bolus administration or for continuousadministration. Injectable formulations may be prepared, packaged, orsold in unit dosage form, such as in ampules or in multidose containerscontaining a preservative. Formulations for parenteral administrationinclude, but are not limited to, suspensions, solutions, emulsions inoily or aqueous vehicles, pastes, and implantable sustained-release orbiodegradable formulations. Such formulations may further comprise oneor more additional ingredients including, but not limited to,suspending, stabilizing, or dispersing agents. In one embodiment of aformulation for parenteral administration, the active ingredient isprovided in dry (i.e., powder or granular) form for reconstitution witha suitable vehicle (e.g., sterile pyrogen-free water) prior toparenteral administration of the reconstituted composition.

The pharmaceutical compositions may be prepared, packaged, or sold inthe form of a sterile injectable aqueous or oily suspension or solution.This suspension or solution may be formulated according to the knownart, and may comprise, in addition to the active ingredient, additionalingredients such as the dispersing agents, wetting agents, or suspendingagents described herein. Such sterile injectable formulations may beprepared using a non-toxic parenterally-acceptable diluent or solvent,such as water or 1,3-butanediol, for example. Other acceptable diluentsand solvents include, but are not limited to, Ringer's solution,isotonic sodium chloride solution, and fixed oils such as syntheticmono- or di-glycerides. Other parentally-administrable formulationswhich are useful include those which comprise the active ingredient inmicrocrystalline form, in a liposomal preparation, or as a component ofa biodegradable polymer system. Compositions for sustained release orimplantation may comprise pharmaceutically acceptable polymeric orhydrophobic materials such as an emulsion, an ion exchange resin, asparingly soluble polymer, or a sparingly soluble salt.

Topical Administration

An obstacle for topical administration of pharmaceuticals is the stratumcorneum layer of the epidermis. The stratum corneum is a highlyresistant layer comprised of protein, cholesterol, sphingolipids, freefatty acids and various other lipids, and includes cornified and livingcells. One of the factors that limit the penetration rate (flux) of acompound through the stratum corneum is the amount of the activesubstance that can be loaded or applied onto the skin surface. Thegreater the amount of active substance which is applied per unit of areaof the skin, the greater the concentration gradient between the skinsurface and the lower layers of the skin, and in turn the greater thediffusion force of the active substance through the skin. Therefore, aformulation containing a greater concentration of the active substanceis more likely to result in penetration of the active substance throughthe skin, and more of it, and at a more consistent rate, than aformulation having a lesser concentration, all other things being equal.

Formulations suitable for topical administration include, but are notlimited to, liquid or semi-liquid preparations such as liniments,lotions, oil-in-water or water-in-oil emulsions such as creams,ointments or pastes, and solutions or suspensions. Topicallyadministrable formulations may, for example, comprise from about 1% toabout 10% (w/w) active ingredient, although the concentration of theactive ingredient may be as high as the solubility limit of the activeingredient in the solvent. Formulations for topical administration mayfurther comprise one or more of the additional ingredients describedherein.

Enhancers of permeation may be used. These materials increase the rateof penetration of drugs across the skin. Typical enhancers in the artinclude ethanol, glycerol monolaurate, PGML (polyethylene glycolmonolaurate), dimethylsulfoxide, and the like. Other enhancers includeoleic acid, oleyl alcohol, ethoxydiglycol, laurocapram, alkanecarboxylicacids, dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone.

One acceptable vehicle for topical delivery of some of the compositionsof the invention may contain liposomes. The composition of the liposomesand their use are known in the art (for example, see Constanza, U.S.Pat. No. 6,323,219).

In alternative embodiments, the topically active pharmaceuticalcomposition may be optionally combined with other ingredients such asadjuvants, anti-oxidants, chelating agents, surfactants, foaming agents,wetting agents, emulsifying agents, viscosifiers, buffering agents,preservatives, and the like. In another embodiment, a permeation orpenetration enhancer is included in the composition and is effective inimproving the percutaneous penetration of the active ingredient into andthrough the stratum corneum with respect to a composition lacking thepermeation enhancer. Various permeation enhancers, including oleic acid,oleyl alcohol, ethoxydiglycol, laurocapram, alkanecarboxylic acids,dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone, are known tothose of skill in the art. In another aspect, the composition mayfurther comprise a hydrotropic agent, which functions to increasedisorder in the structure of the stratum corneum. and thus allowsincreased transport across the stratum corneum. Various hydrotropicagents such as isopropyl alcohol, propylene glycol, or sodium xylenesulfonate, are known to those of skill in the art.

The topically active pharmaceutical composition should be applied in anamount effective to affect desired changes. As used herein “amounteffective” shall mean an amount sufficient to cover the region of skinsurface where a change is desired. An active compound should be presentin the amount of from about 0.0001% to about 15% by weight volume of thecomposition. In particular, it is advantageously present in an amountfrom about 0.0005% to about 5% of the composition; more particularly, itis advantageoulsy present in an amount of from about 0.001% to about 1%of the composition. Such compounds may be synthetically-or naturallyderived.

Buccal Administration

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for buccal administration. Suchformulations may, for example, be in the form of tablets or lozengesmade using conventional methods, and may contain, for example, 0.1 to20% (w/w) of the active ingredient, the balance comprising an orallydissolvable or degradable composition and, optionally, one or more ofthe additional ingredients described herein. Alternately, formulationssuitable for buccal administration may comprise a powder or anaerosolized or atomized solution or suspension comprising the activeingredient. Such powdered, aerosolized, or aerosolized formulations,when dispersed, preferably have an average particle or droplet size inthe range from about 0.1 to about 200 nanometers, and may furthercomprise one or more of the additional ingredients described herein. Theexamples of formulations described herein are not exhaustive and it isunderstood that the invention includes additional modifications of theseand other formulations not described herein, but which are known tothose of skill in the art.

Rectal Administration

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for rectal administration. Such acomposition may be in the form of, for example, a suppository, aretention enema preparation, and a solution for rectal or colonicirrigation.

Suppository formulations may be made by combining the active ingredientwith a non-irritating pharmaceutically acceptable excipient which issolid at ordinary room temperature (i.e., about 20° C.) and which isliquid at the rectal temperature of the subject (i.e., about 37° C. in ahealthy human). Suitable pharmaceutically acceptable excipients include,but are not limited to, cocoa butter, polyethylene glycols, and variousglycerides. Suppository formulations may further comprise variousadditional ingredients including, but not limited to, antioxidants, andpreservatives.

Retention enema preparations or solutions for rectal or colonicirrigation may be made by combining the active ingredient with apharmaceutically acceptable liquid carrier. As is well known in the art,enema preparations may be administered using, and may be packagedwithin, a delivery device adapted to the rectal anatomy of the subject.Enema preparations may further comprise various additional ingredientsincluding, but not limited to, antioxidants, and preservatives.

Additional Administration Forms

Additional dosage forms of this invention include dosage forms asdescribed in U.S. Pat. Nos. 6,340,475; 6,488,962; 6,451,808; 5,972,389;5,582,837; and 5,007,790. Additional dosage forms of this invention alsoinclude dosage forms as described in U.S. Patent Applications Nos.20030147952; 20030104062; 20030104053; 20030044466; 20030039688; and20020051820. Additional dosage forms of this invention also includedosage forms as described in PCT Applications Nos. WO 03/35041; WO03/35040; WO 03/35029; WO 03/35177; WO 03/35039; WO 02/96404; WO02/32416; WO 01/97783; WO 01/56544; WO 01/32217; WO 98/55107; WO98/11879; WO 97/47285; WO 93/18755; and WO 90/11757.

Controlled Release Formulations and Drug Delivery Systems

In one embodiment, the formulations of the present invention may be, butare not limited to, short-term, rapid-offset, as well as controlled, forexample, sustained release, delayed release and pulsatile releaseformulations.

The term sustained release is used in its conventional sense to refer toa drug formulation that provides for gradual release of a drug over anextended period of time, and that may, although not necessarily, resultin substantially constant blood levels of a drug over an extended timeperiod. The period of time may be as long as a month or more and shouldbe a release which is longer that the same amount of agent administeredin bolus form.

For sustained release, the compounds may be formulated with a suitablepolymer or hydrophobic material which provides sustained releaseproperties to the compounds. As such, the compounds for use the methodof the invention may be administered in the form of microparticles, forexample, by injection or in the form of wafers or discs by implantation.

In one embodiment of the invention, the compounds of the invention areadministered to a patient, alone or in combination with anotherpharmaceutical agent, using a sustained release formulation.

The term delayed release is used herein in its conventional sense torefer to a drug formulation that provides for an initial release of thedrug after some delay following drug administration and that may,although not necessarily, includes a delay of from about 10 minutes upto about 12 hours.

The term pulsatile release is used herein in its conventional sense torefer to a drug formulation that provides release of the drug in such away as to produce pulsed plasma profiles of the drug after drugadministration.

The term immediate release is used in its conventional sense to refer toa drug formulation that provides for release of the drug immediatelyafter drug administration.

As used herein, short-term refers to any period of time up to andincluding about 8 hours, about 7 hours, about 6 hours, about 5 hours,about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40minutes, about 20 minutes, or about 10 minutes and any or all whole orpartial increments thereof after drug administration after drugadministration.

As used herein, rapid-offset refers to any period of time up to andincluding about 8 hours, about 7 hours, about 6 hours, about 5 hours,about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40minutes, about 20 minutes, or about 10 minutes, and any and all whole orpartial increments thereof after drug administration.

Dosing

The therapeutically effective amount or dose of a compound of thepresent invention depends on the age, sex and weight of the patient, thecurrent medical condition of the patient and the progression of adisease or disorder contemplated in the invention. The skilled artisanis able to determine appropriate dosages depending on these and otherfactors.

A suitable dose of a compound of the present invention may be in therange of from about 0.01 mg to about 5,000 mg per day, such as fromabout 0.1 mg to about 1,000 mg, for example, from about 1 mg to about500 mg, such as about 5 mg to about 250 mg per day. The dose may beadministered in a single dosage or in multiple dosages, for example from1 to 4 or more times per day. When multiple dosages are used, the amountof each dosage may be the same or different. For example, a dose of 1 mgper day may be administered as two 0.5 mg doses, with about a 12-hourinterval between doses.

It is understood that the amount of compound dosed per day may beadministered, in non-limiting examples, every day, every other day,every 2 days, every 3 days, every 4 days, or every 5 days. For example,with every other day administration, a 5 mg per day dose may beinitiated on Monday with a first subsequent 5 mg per day doseadministered on Wednesday, a second subsequent 5 mg per day doseadministered on Friday, and so on.

In the case wherein the patient's status does improve, upon the doctor'sdiscretion the administration of the inhibitor of the invention isoptionally given continuously; alternatively, the dose of drug beingadministered is temporarily reduced or temporarily suspended for acertain length of time (i.e., a “drug holiday”). The length of the drugholiday optionally varies between 2 days and 1 year, including by way ofexample only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days,12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days,120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days,320 days, 350 days, or 365 days. The dose reduction during a drugholiday includes from 10%-100%, including, by way of example only, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, or 100%.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, the dosage or thefrequency of administration, or both, is reduced, as a function of thedisease or disorder, to a level at which the improved disease isretained. In one embodiment, patients require intermittent treatment ona long-term basis upon any recurrence of symptoms and/or infection.

The compounds for use in the method of the invention may be formulatedin unit dosage form. The term “unit dosage form” refers to physicallydiscrete units suitable as unitary dosage for patients undergoingtreatment, with each unit containing a predetermined quantity of activematerial calculated to produce the desired therapeutic effect,optionally in association with a suitable pharmaceutical carrier. Theunit dosage form may be for a single daily dose or one of multiple dailydoses (e.g., about 1 to 4 or more times per day). When multiple dailydoses are used, the unit dosage form may be the same or different foreach dose.

Toxicity and therapeutic efficacy of such therapeutic regimens areoptionally determined in cell cultures or experimental animals,including, but not limited to, the determination of the LD₅₀ (the doselethal to 50% of the population) and the ED₅₀ (the dose therapeuticallyeffective in 50% of the population). The dose ratio between the toxicand therapeutic effects is the therapeutic index, which is expressed asthe ratio between LD₅₀ and ED₅₀. The data obtained from cell cultureassays and animal studies are optionally used in formulating a range ofdosage for use in human. The dosage of such compounds liesadvantageously within a range of circulating concentrations that includethe ED₅₀ with minimal toxicity. The dosage optionally varies within thisrange depending upon the dosage form employed and the route ofadministration utilized.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures, embodiments, claims, and examples described herein.Such equivalents were considered to be within the scope of thisinvention and covered by the claims appended hereto. For example, itshould be understood, that modifications in reaction conditions,including but not limited to reaction times, reaction size/volume, andexperimental reagents, such as solvents, catalysts, pressures,atmospheric conditions, e.g., nitrogen atmosphere, andreducing/oxidizing agents, with art-recognized alternatives and using nomore than routine experimentation, are within the scope of the presentapplication.

It is to be understood that wherever values and ranges are providedherein, all values and ranges encompassed by these values and ranges,are meant to be encompassed within the scope of the present invention.Moreover, all values that fall within these ranges, as well as the upperor lower limits of a range of values, are also contemplated by thepresent application.

The following examples further illustrate aspects of the presentinvention. However, they are in no way a limitation of the teachings ordisclosure of the present invention as set forth herein.

EXAMPLES

The invention is now described with reference to the following Examples.These Examples are provided for the purpose of illustration only and theinvention should in no way be construed as being limited to theseExamples, but rather should be construed to encompass any and allvariations which become evident as a result of the teaching providedherein.

Example 1: Synthesis

To a solution of commercially available 7 dehydrocholesterol (7DHC) (0.2millimoles in ethyl acetate) and was added 1,2,4-triazolinedione (0 22millimoles) under nitrogen atmosphere, and the system was stirred underthe dark at 0-4 ° C. for 3 hours. The pink color eventually disappeared.The solvent was removed under vacuum.

The residue crude dry residue (0.07 millimoles) was added to a stirredsuspension of bromoacetic acid and dicyclohexylcarbodiimide (DCC) indichloromethane at 0-4 ° C. under a nitrogen gas atmosphere. Thereaction mixture was stirred overnight, filtered to remove the resultingdicyclohexyl urea. The clear solution was evaporated and concentratedunder reduced pressure to generate an oily residue, which was purifiedby preparative TLC.

The well resolved band was extracted with 20% methanol indichloromethane, and the compound was isolated by evaporating thesolvent under the reduced pressure, to produce a white to pale yellowsolid (yield: 35-75% depending on the batch).

Example 2: Reduction of Cell Viability of Ovarian Cancer Cells

Exemplary compounds of the invention were tested in an ovarian cancercell line (SKOV-3), using 7-dehydrocholesterol (7DHC) as a control.

SKOV-3 cells were seeded (5,000 cells/well) in complete DMEM(supplemented with 10% fetal bovine serum and 1% antibiotic) and allowedto adhere overnight. Media was replaced with a fresh complete DMEM mediacontaining varying concentrations of the drugs of interest. DMSO wasused as control. At a suitable interval (varying from 1 minute to 4days), the media was replaced again with complete RPMI (supplementedwith 10% fetal bovine serum and 1% antibiotic) containing MTS dye(Invitrogen Inc.) and incubated for a period varying from 10 minutes to1 day. The absorbance of the media was read at varying wavelengths,advantageously at 492 nM in an ELISA reader.

FIG. 1 illustrates the reduction in cell viability of the SKOV-3 cellsupon treatment with Compound (2).

FIG. 2 illustrates the reduction in cell viability of the SKOV-3 cellsupon treatment with Compound (3).

Example 3: Reduction of Cell Viability of Endometrial Cancer Cells

An exemplary compound of the invention (Compound (3)) was tested in anendometrial cancer cell line (ECC-1).

ECC-1 cells were seeded (10,000 cells/well) in complete RPMI(supplemented with 10% fetal bovine serum and 1% antibiotic) and allowedto adhere over night. Media was replaced with a fresh complete RPMImedia containing varying concentrations of Compound (3). DMSO was usedas control. At suitable intervals (varying from 1 minute to 4 days), themedia was replaced again with complete RPMI (supplemented with 10% fetalbovine serum and 1% antibiotic) containing MTS dye (Invitrogen Inc) andincubated for period (varying from 10 min to 1 day). The absorbance ofthe media was read at varying wavelengths, advantageously at 492 nM inan ELISA reader.

FIG. 3 illustrates the reduction in cell viability of the ECC-1 cellsupon treatment with Compound (3).

Example 4: Inhibition of Lipid Synthesis in Cancer Cells

Lipid synthesis in SKOV-3 cells treated with an exemplary compound ofthe invention was evaluated.

Approximately 3 million cells in serum free conditions were treated withCompound (3) (500 nM). Media was removed, and cells were washed withPBST, trypisinized and collected after centrifugation at 1,000 rpm. Thecell pellet was washed with PBST and treated with methanolic HC1 for 1hour. The methanol was removed, and cells were suspended in water andextracted with chloroform. The collected chloroform layer was evaporatedwith a gentle nitrogen stream, and the residue was analyzed by HPLC. Asillustrated in FIGS. 6A-6B, the lipids appearing at 10.06 minutes weresignificantly reduced upon the treatment with Compound (3), indicatingthat lipid synthesis had been inhibited by the compound of theinvention.

Example 5: Xenographs

The effects of exemplary compounds of the invention on the growth ofSKOV-3 tumor xenographs in nude mice were investigated. Substantively,four- to six week-old immunodeficient nude mice (NU/NU; strain code088/homozygous) (Charles River Laboratories, Wilmington, Mass.) weremaintained at a temperature of 22±1° C. and a relative humidity of55±5%, with a 12 h light/dark cycle. SKOV-3 cells were cultured to 80%confluence, washed in PBS twice, harvested by hypsination, pooled incomplete medium, washed in PBS twice, and 2×10⁶ cells/inoculate weresuspended in 0.1 ml of matrigel and inoculated subcutaneously in theflank of mice. Mice with developing tumors after two weeks were randomlyassigned to experimental groups. Compound (3) was prepared as a stocksolution of 1 mM in 100% EtOH and diluted 1:40 in PBS foradministration. Mice were treated intraperiotoncally every other daywith either vehicle control (control group; 7 animals) or 300 μl (10mg/kg bwt) of MT19c (n=7) for 40 days. Mice were weighed and tumor sizecaculated using a caliper every 5 days.

FIG. 7 illustrates the finding that Compound (3) reduces the tumorburden in one such xenograph model.

Example 6: Reduction of Cell Viability of a Panel of Cancer Cell Lines

Cell lines derived from ovarian cancer, endometrial cancer, breastcancer, prostate cancer and neuroblastoma (5,000 cells/well) were seededin complete DMEM (supplemented with 10% fetal bovine serum and 1%antibiotic) and allowed to adhere over night. Media was replaced with afresh complete DMEM media containing varying concentrations of Compound(3). DMSO was used as control.

At suitable intervals (varying from 1 minute to 4 days), the media wasreplaced again with complete RPMI (supplemented with 10% fetal bovineserum and 1% antibiotic) containing MTS dye (Invitrogen Inc) andincubated for period (varying from 10 min to 1 day). The absorbance ofthe media was read at varying wavelengths, advantageously at 492 nm, inan ELISA reader

Example 7: Ascorbic Acid Pretreatment in a Panel of Cancer Cell Lines

Cell lines derived from ovarian cancer, endometrial cancer, breastcancer, prostate cancer and neuroblastoma (5,000 cells/well) were seededin complete DMEM (supplemented with 10% fetal bovine serum and 1%antibiotic) and allowed to adhere over night. Cells were pretreated with100 millimoles of acrorbic acid in 50 μL DMEM media for 3 hours. To theexisting media was added DMEM media containing varying concentrations of2× concentration of Compound (3). DMSO was used as control. At suitableintervals (varying from 1 minute to 4 days), the media was replacedagain with complete RPMI (supplemented with 10% fetal bovine serum and1% antibiotic) containing MTS dye (Invitrogen Inc) and incubated forperiod (varying from 10 min to 1 day). The absorbance of the media wasread at varying wavelengths, advantageously at 492 nm, in an ELISAreader. As demonstrated, pretreatment with ascorbioc acid reduced thecytotoxic effects of Compound (3) in a panel of cancer cell lines.

Example 8: Inhibition of Cell Migration in a Wound Healing Assay

A 24-well plate was coated with 100 L of sterile 1% gelatin solution perwell. After gelatin coating was hardened, about 100,000 HUVEC cells wereseeded per well and allowed to grow to 100% confluence in EBM-2 media. A200 L pipette tip was then used to create a vertical scratch wound downthe center of each well, and cells were washed once with fresh EBM-2media. Cell monolayers were then imaged at a 0 h time point using lightmicroscopy on an inverted microscope under sterile conditions. Woundareas were imaged again using light microscopy, and areas weredemarcated on representative images.

Floating cells were removed and media was replaced with HUVEC cell mediasupplemented with vehicle or Compound (3) (500 nMoles) and incubated for24 hours. The cells were imaged using a pre-cooled inverted NICONmicroscope (40×). The vehicle treated cells recovered the scratched areawhereas compound (3)- treated cells failed to recover and repopulate thescratched area.

Example 9: Inhibition of Microcapillary Formation in a Rat Aorta RingAssay

Rat aorta were harvested from the freshly euthanized naïve rats, cleanedand cut into fine rings and placed on matrigel beds prepared 30 minearlier in a 6 well plate. The aorta were allowed to stay there for 10days, by which time microcapillaries were found to be sprouting. Themedia was replaced with fresh supplemented media and allowed theincubation for next 48 hours, thus affording a strong network of themicrocappilary network. The media was replaced with fresh mediacontaining vehicle or compound concentrations (50 nmoles and 100 nmoles)and incubated for 48 hours. The phase contrast images were recorded on apre-cooled inverted NICON microscope.

Example 10: Antagonism of the Vitamin D Nuclear Receptor (VDR)

The assay was described in detail in Brard et al., 2011, GynecologicOncology. The biochemical interactions between MeTC7, its precursorssuch as 7-DHC or its adduct, and VDR were investigated using afluorescence polarization assay. MeTC7 was incubated with VDR-LBD and afluorescent labeled coactivator peptide (SRC2-3) in the presence andabsence of calcitriol. In the presence of calcitriol, VDR interacts withthe coactivator peptide SRC2-3. VDR antagonists disrupt this interactionby a direct or allosteric mode of inhibition.

MeTC7 showed antagonistic effect at a concentration of 1.5-3 M andhigher (FIGS. 10A-10C). The ability of MeTC7 to bind to VDR and initiatethe conformational change of VDR to allow coactivator recruitment wasdetermined in the absence of calcitriol. Results show that MeTC7 is nota VDR agonist (FIG. 10B). FIG. 10D is a graph that illustrates resultsfor the VDR transactivation assay (as % effect vs. log [Conc]).

Determination of Agonistic/Antagonistic Properties of MeTC7 using aPPAR—Coactivator Binding Assay (FIGS. 10E-10F)

Briefly, pET15b-PPAR -LBD expression plasmid, encoding the PPAR -LBD(amino acids 173-475) was a generous gift from Gabor J. Tigyi(University of Tennessee, Memphis). PPAR-LBD was expressed in BL21 (DE3)(Tnvitrogen), purified by affinity chromatography, and stored at −80° C.in buffer (50 mM Tris (pH 8.0), 25 mM KCl, 2 mM DTT, 10% glycerol, 0.01%NP-40). For the assay, MeTC7 or its precursors were serially diluted inDMSO and 100 nl of each concentration was transferred into 20 μL proteinbuffer (20 mM TRIS (pH 7.5), 100 mM NaCl, 0.01% NP-40, 2% DMSO, 10 nMDRIP2 (CNTKNHPMLMNLLKDNPAQD) labeled with Texas-Red maleimide, and 1 MPPAR-LBD) in the presence and absence of rosiglitazone (5 M) inquadruplet using black 384 well plate (Costar, #3658). The samples wereallowed to equilibrate for two hours. Binding was then measured usingfluorescence polarization (excitation 595 nm, emission 615 nm) using aM1000 plate reader (Tecan). The experiments were evaluated usingGraphPad Prism 5, and IC50 values were obtained by fitting the data toan equation (Sigmoidal dose-response-variable slope (four parameters).Values are given as the mean values of two independent experiments witha 95% confidence interval.

Example 11: Further Biological Characterization FIGS. 11A-11C:

Viability of cell lines (ECC-1, AN3CA, RL-95-2, SKOV-3, PC-3, MCF-7,SMSKCNR and TCL-1) before and after MeTC7 treatment was determined bythe CellTiter 96® AQueous One Solution Assay (Promega Corp, Madison,Wisc.) following the manufacturer's recommendations. This colorimetricassay is based on the ability of mitochondria to reduce a substrate[MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] into a soluble formazanproduct quantified by measuring the absorbance at 490 nm. The resultingOD is directly proportional to the number of living cells. Briefly,cells (5×10³/well) were plated into 96 well flat bottom plates (Corning,Inc., Coming, N.Y.) before treatment with various drugs or vehicle(DMSO) as indicated. Following incubation at 37° C. in a cell cultureincubator for 20 h MTS reagent was added at a 1:10 dilution to themedium. The samples were incubated for an additional 4 h beforeabsorbance was measured at 490 nm in an ELISA plate reader (ThermoLabsystems, Waltham, Mass.). Experiments were performed in triplicates;data are expressed as the mean of the triplicate determinations (X±SD)of a representative experiment in % of absorbance by samples withuntreated cells [=100%]. Similarly, the cell viability of HEPG2 andHEK293 cells was determined except that the drug concentrations wereused in the logarithmic values.

FIG. 11D:

SKOV -3 and OVCAR-8 ovarian cancer cells (10,000 per well) were seededin a 8 well slide chamber in DMEM complete media and allowed to incubateovernight. Cells were treated with complete media containing eithervehicle or MeTC7 (500 nM) and incubated for 12 hours in a humidifiedincubator maintained at standard conditions. DNA fragmentation wasdetected using the DeadEndTM Fluorometric TUNEL System assay (Promega,Madison, Wisc.) according to the manufacturer's recommendations.Fluorescence of apoptotic cells (green; labeling of DNA nicks byfluorescein-12-dUTP) and of chromatin (red; staining of chromatin withpropidium iodide) was detected by fluorescence microscopy with aninverted microscope (Nikon Eclipse TE2000-E) and a 10× objective. Fourrandomly chosen microscopic fields were captured. For the detection ofthe cytochrome-C release as a marker of the apoptosis, the cells werefixed in 5% cold formalin solution and cells were stained withcytochrome-C primary antibody (Santa Cruz Biotechnology, USA) andcorresponding fluroescence linked secondary antibody. The images wererecorded using an inverted microscope. At least five field werearbitrarily scoped. A representative field is shown with abundantcytochrome-C release in the cytosol in the MeTC7 treated cells comparedto the control which shows the concentrated staining of cytochrome-C inthe nucleus.

FIG. 11E:

SKOV-3 and OVCAR-8 ovarian cancer cells were purchased from AmericanTissue Culture Collection (ATCC) (www dot atcc dot org) and maintainedin DMEM media (Invitrogen Inc) supplemented with fetal bovine serum(10%) and antibiotics (1%). SKOV-3 cells (1 million each) were seeded ina 100mm² petri dishes containing 5 mL of the complete DMEM media andcells were allowed to adhere and incubate overnight. Media was replacedwith media containing vehicle or MeTC7 (500 nM) 12 hours. Media wascollected and stored at −20° C. for future studies. Preparation of celllysates. PAGE and immunoblotting with appropriate antibodies purchasedfrom Origene (MD, USA) was carried out as previously described (Moore etal., Plos One, 2012, DOI: 10.1371journal dot pone dot 0034443). Briefly,protein concentration of the remaining supernatant of the cell lysatewas quantitated (BioRad protein estimation kit, Hercules, Calif.) andWestern blotting was carried out. Samples were boiled in the presence of5× SDS-PAGE sample buffer and 50 g total cellular protein/lane wereseparated on 12% SDS-polyacrylamide gels and blotted onto PVDFmembranes. The blots were blocked with 5% nonfat dry milk in PBST for 1hr at room temperature and incubated overnight at 4° C. with theantibodies against caspase-7, 8 and cleaved PARP-1. After washing inPBST the blots were incubated with secondary antibody(peroxidase-conjugated antibodies; Amersham-Pharmacia Biotech,Piscataway, N.J.). The bands were visualized using horseradishperoxidase-conjugated secondary antibodies (Amersham-Pharmacia Biotech,Piscataway, N.J.) and documented by autoradiography (Fast Film, Phenix,Hayward, Calif.).

FIGS. 11F-11H:

Animal experiments were carried out in the animal facilities of RhodeIsland Hospital (RIH), Rhode Island, USA with strict adherence to theguidelines of the Animal Welfare Committee of Rhode Island Hospital(RIH) and Women and Infants Hospital of Rhode Island (Laboratory AnimalProtection Approval: A3922-01). Four to six week-old immunodeficientnude mice (NU/NU; strain code 088/homozygous) (Charles RiverLaboratories, Wilmington, Mass.) were maintained at a temperature of−22° C. and a relative humidity of −55%, with a 12 h light/dark cycle.SKOV-3 cells were cultured to 80% confluence, washed in PBS twice,harvested by trypsination, pooled in complete medium, washed in PBStwice, and 1×10⁶ cells/inoculate were suspended in 0.1 ml of matrigeland inoculated subcutaneously in the flank of mice. Mice with developingtumors after two weeks were randomly assigned to experimental groups.Mice were treated intraperitoneally every day with either vehiclecontrol (control group; 6 animals) or MeTC7 (n=6, 10 mg/kg bwt, 5× week)for 31 days. Mice were weighed and tumor size were measured using adigital caliper on the days as indicated in the FIGS. 11F-11H. Thesurvival curve of the animals survived during the course of the animaltrial in treatment group compared to the control group (FIG. 11G) wasestimated by the Kaplan-Meier analysis.

FIG. 11I:

The xenograft tissues from the animals of FIG. 11I were harvested aftereuthanasia and fixed in paraformaldehyde and embedded in paraffin. Theslides of 5 M were stained for the expression of VDR using the primaryantibody (Santa Cruz Biotechnology, 1:50 diluation, USA.) andcorresponding flourescence linked secondary and images were recorded asdescribed previously (Moore et al., Plos One, 2012, DOI: 10.1371/journaldot pone dot 0034443).

Example 12: Medulloblastoma Xenograft

Animal experiments were carried out in the animal facilities of RhodeIsland Hospital (RIH), Rhode Island, USA with strict adherence to theguidelines of the Animal Welfare Committee of Rhode Island Hospital(RIH) and Women and Infants Hospital of Rhode Island (Laboratory AnimalProtection Approval: A3922-01). Four to six week-old immunodeficientnude mice (NU/NU; strain code 088/homozygous) (Charles RiverLaboratories, Wilmington, Mass.) were maintained at a temperature of˜22° C. and a relative humidity of ˜55%, with a 12 h light/dark cycle.DAOY cells were cultured to 80% confluence, washed in PBS twice,harvested by trypsination, pooled in complete medium, washed in PBStwice, and 1×10⁶ cells/inoculate were suspended in 0.1 ml of matrigeland inoculated subcutaneously in the flank of mice. Mice with developingtumors after two weeks were randomly assigned to experimental groups.Mice were treated intraperitoneally every day with either vehiclecontrol (control group; 6 animals) or MeTC7 (n=5, 10 mg/kg bwt, 5× week)for 15 days. Mice were weighed and tumor size were measured using adigital caliper every 5^(th) days.

Example 13: Melanoma Xenograft

Animal experiments were carried out in the animal facilities of RhodeIsland Hospital (RIH), Rhode Island, USA with strict adherence to theguidelines of the Animal Welfare Committee of Rhode Island Hospital(RIH) and Women and Infants Hospital of Rhode Island (Laboratory AnimalProtection Approval: A3922-01). Four to six week-old immunodeficientnude mice (NU/ NU; strain code 088/homozygous) (Charles RiverLaboratories, Wilmington, Mass.) were maintained at a temperature of˜22° C. and a relative humidity of ˜55%, with a 12 h light/dark cycle.A2058 cells were cultured to 80% confluence, washed in PBS twice,harvested by trypsination, pooled in complete medium, washed in PBStwice, and 1×10⁶ cells/inoculate were suspended in 0.1 ml of matrigeland inoculated subcutaneously in the flank of mice. Mice with developingtumors after two weeks were randomly assigned to experimental groups.Mice were treated intraperitoneally every day with either vehiclecontrol (control group; 7 animals) or MeTC7 (10 mg/kg bwt, 5× week) for15 days. Mice were weighed and tumor size were measured using a digitalcaliper every 5^(th) days. The percent of animals with tumor size lessthan 15 mm was calculated by Kaplan-Meier Analysis.

The disclosures of each and every patent, patent application, andpublication cited herein are hereby incorporated herein by reference intheir entirety.

While this invention has been disclosed with reference to specificembodiments, it is apparent that other embodiments and variations ofthis invention may be devised by others skilled in the art withoutdeparting from the true spirit and scope of the invention. The appendedclaims are intended to be construed to include all such embodiments andequivalent variations.

1.-71. (canceled)
 72. A method of treating an immunogenic cancer in asubject in need thereof, the method comprising administering to thesubject a therapeutically effective amount of at least one compound offormula (I) or a salt or solvate thereof: wherein:

R₁ is CR₅ or N; R₃ is selected from the group consisting of—O(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and—OC(═O)C(R₅)=C(R₅)₂; R₂ is selected from the group consisting of O, S,C(R₄)₂, and N(R₄); each occurrence of R₄ is independently selected fromthe group consisting of H, alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl,substituted heteroarylalkyl, OR₅, and N(R₅)₂; each occurrence of Rs isindependently selected from the group consisting of H, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, arylalkyl, substituted arylalkyl, heteroaryl, substitutedheteroaryl, heteroarylalkyl, and substituted heteroarylalkyl; R₆ isselected from the group consisting of F, Cl, Br, I, mesyl, tosyl,—OSi(R₅)₃, —C(═O)OR₅, and —C(═O)R₅; the dotted line is a single ordouble bond; and n is an integer ranging from 1 to
 10. 73. The method ofclaim 72, wherein the compound of formula (I) is the compound of formula(Ia), or a salt or solvate thereof:


74. The method of claim 72, wherein the compound of formula (I) is thecompound of formula (Ib), or a salt or solvate thereof:


75. The method of claim 72, wherein R₁ is N.
 76. The method of claim 75,wherein R₂ is N(R₄).
 77. The method of claim 76, wherein the compound offormula (I) is the compound of formula (Ic), or a salt or solvatethereof:


78. The method of claim 77, wherein the compound of formula (I) is thecompound of formula (Id), or a salt or solvate thereof:


79. The method of claim 72, wherein the compound of formula (I) isselected from the group consisting of


80. The method of claim 72, wherein the immunogenic cancer comprises acancer selected from the group consisting of malignant melanoma, renalcell carcinoma, Mantel cell lymphoma, follicular lymphoma, diffuse largeB-cell lymphoma, T-cell acute lymphoblastic leukemia, Burkitt lymphoma,myeloma, immunocytoma, acute promyelocytic leukemia, chronicmyeloid/acute lymphoblastic leukemia, acute leukemia, B-cell acutelymphoblastic leukemia, anaplastic large cell leukemia, myelodysplasticsyndrome/acute myeloid leukemia, non-Hodgkin's lymphoma, chroniclymphocytic leukemia, acute myelogenous leukemia (AML), common (pre-B)acute lymphocytic leukemia, malignant melanoma, T-cell lymphoma,leukemia, B-cell lymphoma, epithelial malignancies, lymphoidmalignancies, gynecologic carcinoma, biliary adenocarcinoma, and ductaladenocarcinoma of the pancreas.
 81. The method of claim 72, wherein thesubject is further administered at least one additional chemotherapeuticagent.
 82. The method of claim 81, wherein the at least one additionalchemotherapeutic agent is selected from the group consisting ofalkylating agents; nitrosoureas; antimetabolites; antitumor antibiotics;plant alkyloids; taxanes; hormonal agents; antiangiogenesis agents,bleomycin, hydroxyurea, L-asparaginase, and procarbazine.
 83. The methodof claim 81, wherein the compound of formula (I) and the at least oneadditional chemotherapeutic agent are separately administered to thesubject.
 84. The method of claim 81, wherein the compound of formula (I)and the at least one additional chemotherapeutic agent areco-administered to the subject.
 85. The method of claim 72, wherein thecompound of formula (I) is administered to the subject by at least oneroute selected from the group consisting of intravenous, oral,inhalational, rectal, vaginal, transdermal, intranasal, buccal,sublingual, parenteral, intrathecal, intragastrical, ophthalmic,pulmonary and topical routes.
 86. The method of claim 72, wherein thesubject is a mammal.
 87. The method of claim 86, wherein the mammal is ahuman.
 88. The method of claim 72, wherein the subject is furtheradministered radiation therapy.
 89. A method of inhibiting angiogenesisin a subject in need thereof, the method comprising administering to thesubject a therapeutically effective amount of at least one compound offormula (I) or a salt or solvate thereof:

wherein: R₁ is CR₅ or N; R₃ is selected from the group consisting of—O(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)R₆, —OC(═O)(CR₅)_(n)OR₅, and—OC(═O)C(R₅)═C(R₅)₂; R₂ is selected from the group consisting of O, S,C(R₄)₂, and N(R₄); each occurrence of R₄ is independently selected fromthe group consisting of H, alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, aryl, substituted aryl, arylalkyl, substitutedarylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl,substituted heteroarylalkyl, OR₅, and N(R₅)₂; each occurrence of R₅ isindependently selected from the group consisting of H, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, arylalkyl, substituted arylalkyl, heteroaryl, substitutedheteroaryl, heteroarylalkyl, and substituted heteroarylalkyl; R₆ isselected from the group consisting of F, Cl, Br, I, mesyl, tosyl,—OSi(R₅)₃, —C(═O)OR₅, and —C(═O)R₅; the dotted line is a single ordouble bond; and n is an integer ranging from 1 to
 10. 90. The method ofclaim 89, wherein the compound of formula (I) is the compound of formula(Ia), or a salt or solvate thereof:


91. The method of claim 72, wherein the compound of formula (I) is thecompound of formula (Ib), or a salt or solvate thereof:


92. The method of claim 89, wherein R₁ is N.
 93. The method of claim 92,wherein R₂ is N(R₄).
 94. The method of claim 93, wherein the compound offormula (I) is the compound of formula (Ic), or a salt or solvatethereof:


95. The method of claim 94, wherein the compound of formula (I) is thecompound of formula (Id), or a salt or solvate thereof:


96. The method of claim 89, wherein the compound of formula (I) isselected from the group consisting of


97. The method of claim 89, wherein the subject is further administeredat least one additional anti-angiogenesis agent.
 98. The method of claim97, wherein the at least one additional anti-angiogenesis agent isselected from the group consisting of 2-methoxyestradiol AG3340,angiostatin, antithrombin-III, anti-VEGF antibody, VEGF antagonist,batimastat, bevacizumab, BMS-275291, CA1, canstatin, combretastatin,combretastatin-A4 phosphate, CC-5013, captopril, celocoxib, dalteparin,EMD121974, endostatin, erlotinib, gefitinib, genistein, halofuginone,ID1, ID3, IM862, omatinib mesylate, inducible protein-10,interferon-alpha, interleukin-12, lavendustin-a, LY317615, AE-941,merimastat, mapsin, medroxpregesteron acetate, Meth-1, Meth-2,Neovastat, osteopontin cleaved product, PEX, pigment epithelium growthfactor, platelet growth factor 4, prolactin fragment, proliferin-relatedprotein (PRP), PTK787/ZK222584, recombinant human platelet factor-4,restin, squalamine, SU5416, SU6668, suramin, taxol, tecogolan,thalidomide, thrombospondin, TNP-470, troponin I, vasostatin, VEGF1,VEGF-TRAP and ZD6474.
 99. The method of claim 97, wherein the compoundand the at least one additional anti-angiogenesis agent are separatelyadministered to the subject.
 100. The method of claim 97, wherein thecompound and the at least one additional anti-angiogenesis agent areco-administered to the subject.
 101. The method of claim 89, wherein thesubject is a mammal.
 102. The method of claim 101, wherein the mammal isa human.